[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"$fxLN3MUwXCdr5RPjwZYIDpOj8CHyjOmngWTgoKXPtZbg":3,"$fD_Bxq1HcTAGfRZnSoA_haZGQyWSj88_PsWBrJFaWq9M":36,"$f3Ft0rKFJHppdzE-vuveecxx1BUcg9iOlMLtyzf_MJDg":161,"$fucxFBm2ZjZfGSdmdRaSNGBI_F0jJme4f0GTvzUhQfL8":226},[4,8,12,16,20,24,28,32],{"title":5,"slug":6,"path":7},"About Microbeonline.com","about-microbeonline-com","\u002Fabout-microbeonline-com\u002F",{"title":9,"slug":10,"path":11},"About Me","about-me","\u002Fabout-microbeonline-com\u002Fabout-me\u002F",{"title":13,"slug":14,"path":15},"Advertise with Us","advertise-us","\u002Fadvertise-us\u002F",{"title":17,"slug":18,"path":19},"Privacy Policy","privacy-policy","\u002Fprivacy-policy\u002F",{"title":21,"slug":22,"path":23},"Authors","authors","\u002Fauthors\u002F",{"title":25,"slug":26,"path":27},"Microbes","microbes","\u002Fmicrobes\u002F",{"title":29,"slug":30,"path":31},"Books","recommended-books","\u002Frecommended-books\u002F",{"title":33,"slug":34,"path":35},"Tags","tags","\u002Ftags\u002F",{"type":37,"data":38},"blog",{"slug":39,"title":40,"description":41,"seoTitle":42,"seoDescription":42,"author":43,"createdDate":44,"lastUpdatedDate":45,"draft":46,"category":47,"image":42,"body":48,"faq":49,"commentsClosed":46,"tags":65,"related":67,"comments":157},"nested-pcr-principle-applications","Nested PCR: Principle and Applications","Nested PCR uses two successive PCR reactions with outer and inner primer sets to maximise sensitivity and specificity. Learn its principle, applications, and contamination risk.",null,"Acharya Tankeshwar","2019-12-20","2026-07-05",false,"lab-equipment","A 45-year-old farmer from the Bihar region of India presents with two weeks of fever, headache, and a eschar — a dark, crusted skin lesion at the site of a mite bite on his forearm. The clinician suspects scrub typhus caused by *Orientia tsutsugamushi*. Blood is drawn and sent to the reference laboratory.\n\nStandard PCR on the blood sample returns negative. The organism is present — the clinical picture is clear — but the bacterial load in peripheral blood in scrub typhus is extremely low, often just a few copies per milliliter. Standard PCR lacks the sensitivity to detect it reliably at that concentration.\n\nThe laboratory repeats the test using nested PCR. The outer primer pair amplifies a large fragment of the *Orientia* 47-kDa gene. The product of that first reaction becomes the template for the second reaction, where inner primers amplify a smaller, specific region nested within the first amplicon. The signal is amplified twice over. This time, the result is positive.\n\nNested PCR did not just improve sensitivity here — it made the diagnosis possible at all. In infections where organisms circulate in blood at very low copy numbers, nested PCR is often the only PCR method capable of detection.\n\nNested PCR is a modification of [PCR](\u002Fpolymerase-chain-reaction-pcr-steps-types-applications\u002F) designed to increase the sensitivity and specificity of the assay reaction. It involves the use of two primer sets directed against the same target and two successive PCR reactions.\n\n![ - Nested PCR (Image Source:https:\u002F\u002Fwww.thermofisher.com\u002F)](https:\u002F\u002Fassets.microbeonline.com\u002Fblogs\u002FNested-PCR-Two-Step-Process.jpg)Figure: Nested PCR (Image Source:https:\u002F\u002Fwww.thermofisher.com\u002F)\n\nThe first set of primers is designed to anneal to sequences upstream from the second set of primers, whereas the second set of primers is situated internally or nested with respect to the first set of primers. The first set of primers also called **“outer primers”** amplify a large fragment of the gene which is used as a template in the second round of PCR that targets a smaller region of the amplicon using the second set of primers also known as **“inner primers or nested primers.”**\n\nThe traditional approach to nested PCR was to perform a number of PCR cycles using the first set of primers, and then open the reaction vessel and add the second, nested, set of primers to run the second PCR cycle. The major problem with this approach is amplicon contamination in the laboratory and a consequential loss of specificity of the assay. To address this issue **single-tube nested PCR (STNPCR) reactions** have been developed, wherein both sets of primers are added to the initial reaction vessel and an extended PCR is performed.\n\nAmplicons from this PCR assays are visualized by electrophoresing the reaction mixture in 2% ethidium bromide-stained [agarose gel](\u002Fagarose-gel-electrophoresis\u002F) along with a molecular weight marker.\n\n![ - Nested Polymerase Chain Reaction (PCR)](https:\u002F\u002Fassets.microbeonline.com\u002Fblogs\u002FNested-PCR-Procedure.jpg)Figure: Nested Polymerase Chain Reaction (PCR)\n\n> Nested PCRs are sometimes necessary to compensate for inefficient first-round PCR due to primer mismatches so, if we can use well-matched primers for first-round PCR nested approach may not be needed in many circumstances.\n\n### Advantage\n\n**It reduces the nonspecific amplification of the target sequence.**\n\nThis is because nested primers will not find priming sites on any primer dimers or nonspecific artifacts generated in the primary PCR. Nested primers will only prime any specific product generated in the primary PCR, thus maintaining PCR specificity.\n\n### Applications\n\nTo improve the sensitivity of the assay, it has been used in many PCR assays. It is particularly useful for suboptimal nucleic acid samples, such as those extracted from formalin-fixed, paraffin-embedded tissue\n\nNested PCRs have proven valuable for the detection of microorganisms when they are present in very low quantities. For example:\n\n- detection of *Rickettsia, Bartonella*, and similar organisms in the blood (bacteremia) and tissues,\n- detection of herpesvirus and enterovirus in the CSF, and\n- detection of *M. tuberculosis* in a sputum sample.\n\n**Commercial application**\n\nThe [BioFire FilmArray](https:\u002F\u002Fwww.biofiredx.com\u002Fproducts\u002Ffilmarray\u002F) from bioMérieux is a commercially available system that employs nested, [multiplex](\u002Fmultiplex-pcr-principle-applications-and-limitations\u002F), and singleplex PCR reactions for the detection of a variety of pathogens.\n\nFor an overview of all PCR types used in clinical microbiology, see [Polymerase Chain Reaction: Steps, Types, and Applications](https:\u002F\u002Fmicrobeonline.com\u002Fpolymerase-chain-reaction-pcr-steps-types-applications\u002F)\n\n### Limitations\n\n- **Susceptible to contamination:** The extreme sensitivity of this type of PCR comes with its own set of problems. This test is highly susceptible to contamination as it involves more time for sample manipulation. Contamination mostly occurs during the transfer of the first-round product to the second tube for the second round of amplification.\n- **Costly:** This PCR assay is also more costly as it involves the use of two separate reactions to arrive at one result. The cost will increase dramatically if assays are repeated when contamination occurs.\n\n## How to Remember\n\n**Nested PCR = two rounds of amplification, each inside the previous.** The name \"nested\" is the memory device itself. The inner primers sit *inside* the sequence amplified by the outer primers — like a smaller box nested inside a larger one. Round 1 (outer primers) amplifies a large region. Round 2 (inner primers) amplifies a smaller region *within* that product. Two rounds of amplification mean exponentially more copies of the target — and two rounds of primer specificity mean far fewer non-specific products.\n\n**Outer primers = big net; inner primers = fine sieve.** The outer primers cast a wide net over the target region, amplifying a large fragment — even from a degraded or low-copy template. The inner primers then act as a fine sieve, amplifying only the specific sequence of interest from within that large fragment. Anything non-specific that slipped through the first net is unlikely to contain the inner primer binding sites and is eliminated in the second round.\n\n**High sensitivity comes with high contamination risk — the trade-off.** Nested PCR is the most sensitive PCR method for low-copy targets — and also the most contamination-prone, because the first-round product is opened and transferred to a second tube in traditional two-tube nested PCR. Even nanogram quantities of that first-round amplicon contaminating the laboratory environment can seed false positives in subsequent runs. This is why single-tube nested PCR (STNPCR) was developed — both primer sets added at the start, no tube opening between rounds.\n\n**Clinical trigger for nested PCR: \"present in very low quantities.\"** Whenever a question describes an organism present in very low copy numbers — *Rickettsia*, *Bartonella*, *M. tuberculosis* in paucibacillary samples, viruses in CSF, *Leishmania* in tissue — nested PCR is the answer. The exam pattern is: low pathogen load + need for high sensitivity = nested PCR.\n\n## Key exam facts in one table\n\n| Topic | Key fact |\n| --- | --- |\n| Definition | Two successive PCR reactions using two primer sets targeting the same region |\n| Outer primers | Amplify a large fragment in the first PCR reaction; also called external primers |\n| Inner primers | Amplify a smaller region nested within the first amplicon; also called nested primers |\n| Why two rounds increase sensitivity | First-round product is a concentrated, enriched template for the second round — low-copy targets undetectable in one round become detectable |\n| Why two rounds increase specificity | Inner primers only bind within the specific first-round product; non-specific first-round products rarely contain inner primer binding sites |\n| Traditional nested PCR — contamination risk | Opening the first-round tube to transfer product releases amplicons into the lab environment; causes false positives in subsequent runs |\n| STNPCR | Single-tube nested PCR — both primer sets added to the initial reaction vessel; no tube opening required; reduces contamination risk |\n| Clinical applications | *Rickettsia*, *Bartonella* in blood; herpesvirus and enterovirus in CSF; *M. tuberculosis* paucibacillary samples; *Leishmania* in tissue; formalin-fixed paraffin-embedded samples |\n| Commercial application | BioFire FilmArray — combines nested, multiplex, and singleplex PCR in an automated closed pouch system |\n| Main advantage | Highest sensitivity of any PCR method for low-copy targets |\n| Main limitation | Susceptible to carry-over contamination; higher cost (two reactions per result) |\n| When to choose nested PCR | Organism present in very low quantities; suboptimal nucleic acid quality; when standard PCR sensitivity is insufficient |\n\n### References and further reading\n\n1. Chang-Hui Shen (2019). Amplification of Nucleic Acids. *Diagnostic Molecular Biology*. Academic Press. \u003Chttps:\u002F\u002Fdoi.org\u002F10.1016\u002FB978-0-12-802823-0.00009-2>\n2. Deepachandi, B., Weerasinghe, S., Soysa, P. et al (2019). A highly sensitive modified nested PCR to enhance case detection in leishmaniasis. *BMC Infectious Diseases* 19, 623. \u003Chttps:\u002F\u002Fdoi.org\u002F10.1186\u002Fs12879-019-4180-3>\n3. Souza G., Almeida A., Farias A., Leal N., Abath F. (2007). Development and Evaluation of a Single Tube Nested PCR Based Approach (STNPCR) for the Diagnosis of Plague. In: Perry R.D., Fetherston J.D. (eds) *The Genus Yersinia*. Advances in Experimental Medicine and Biology, vol 603. Springer.",[50,53,56,59,62],{"question":51,"answer":52},"What is nested PCR and how does it increase sensitivity?","Nested PCR uses two successive PCR reactions targeting the same region. The first reaction uses outer primers to amplify a large fragment of the target sequence. The product of that first reaction becomes the template for the second reaction, which uses inner (nested) primers to amplify a smaller region within the first amplicon. This double amplification increases sensitivity dramatically — a target present in too few copies to be reliably detected in a single PCR round is enriched by the first reaction, making it readily detectable in the second. Nested PCR is the most sensitive PCR method for detecting organisms present in very low quantities.",{"question":54,"answer":55},"Why does nested PCR also increase specificity?","Nested PCR increases specificity because the inner primers only find binding sites within the specific first-round product. Non-specific products generated in the first round — amplicons from non-target sequences that share partial homology with the outer primers — are unlikely to contain binding sites for the inner primers. The second round of amplification therefore selectively amplifies the genuine target while eliminating most non-specific products from the first round. Two independent primer-binding events on the same target provide a level of confirmation that a single primer set cannot offer.",{"question":57,"answer":58},"What is single-tube nested PCR (STNPCR) and why was it developed?","Traditional nested PCR requires opening the first-round reaction tube to transfer product to a second tube for the second PCR reaction. This step releases amplicons into the laboratory environment, where they can contaminate subsequent runs and cause false positives — a serious problem given nested PCR's extreme sensitivity. Single-tube nested PCR (STNPCR) was developed to eliminate this risk: both sets of primers are added to the initial reaction vessel before cycling begins, and an extended PCR protocol activates the inner primers at the appropriate stage without ever opening the tube. STNPCR maintains the sensitivity advantages of nested PCR while reducing contamination risk.",{"question":60,"answer":61},"What clinical infections is nested PCR particularly useful for?","Nested PCR is particularly useful for infections where the pathogen circulates in blood or tissues at very low copy numbers, making standard PCR unreliable. Established applications include: detection of Rickettsia and Bartonella in blood during bacteraemia; detection of M. tuberculosis in paucibacillary samples (e.g., CSF in TB meningitis, pleural fluid); detection of herpesvirus and enterovirus in CSF; detection of Leishmania in tissue biopsies; and detection of organisms in formalin-fixed paraffin-embedded tissue where nucleic acid degradation limits PCR sensitivity. The BioFire FilmArray commercial system uses nested PCR as its first amplification stage.",{"question":63,"answer":64},"What are the main limitations of nested PCR?","The two main limitations are contamination risk and cost. Contamination risk is the most serious: the extreme sensitivity that makes nested PCR valuable also makes it exquisitely susceptible to carry-over contamination. First-round amplicons opening into the laboratory environment can contaminate reagents, surfaces, and subsequent reactions, producing false positives that are difficult to identify and eliminate. STNPCR mitigates but does not entirely eliminate this risk. Cost is the second limitation: two separate PCR reactions are required to produce one result, and if contamination causes a repeat, cost escalates further. Nested PCR is therefore reserved for situations where standard PCR lacks sufficient sensitivity rather than used as a first-line method.",[66],"pcr-techniques",[68,96,135],{"slug":69,"title":70,"description":71,"seoTitle":72,"seoDescription":73,"author":43,"createdDate":74,"lastUpdatedDate":75,"draft":46,"category":47,"image":42,"faq":76,"tags":95},"polymerase-chain-reaction-pcr-steps-types-applications","Polymerase Chain Reaction (PCR): Steps, Types, and Applications","PCR amplifies DNA exponentially in three steps: denaturation, annealing, and extension. Learn the components, steps, types: nested, multiplex, real-time, RT-PCR and clinical applications in diagnostic microbiology.","PCR: Steps, Reagents, Result Interpretation, and Applications","Review PCR reagents and the denaturation, annealing, and extension cycle, then compare major PCR variants, controls, interpretation, and applications.","2016-07-07","2026-08-15",[77,80,83,86,89,92],{"question":78,"answer":79},"What is polymerase chain reaction (PCR) and what does it do?","\u003Cp>Polymerase chain reaction (PCR) is an in vitro molecular technique that amplifies a specific DNA or RNA sequence exponentially, producing up to 10 million copies from a single starting template within a few hours. It works by repeatedly cycling through three temperature-controlled steps (denaturation, annealing, and extension), using a heat-stable DNA polymerase (Taq polymerase) and short synthetic primers that define the target sequence. In clinical microbiology, PCR directly detects a pathogen's nucleic acid in a patient specimen, regardless of whether the organism is alive, cultivable, or present in small quantities.\u003C\u002Fp>",{"question":81,"answer":82},"What are the three steps of PCR and what temperature is used for each?","\u003Cp>PCR has three steps that repeat in each cycle. Denaturation occurs at 94–96°C, heat breaks the hydrogen bonds between the two DNA strands, separating them into single-stranded templates. Annealing occurs at 45–65°C, the temperature is lowered so primers can bind to their complementary sequences on each strand. Extension occurs at 72°C, Taq polymerase synthesizes a new complementary DNA strand starting from each primer. After 30–40 cycles, the target sequence is amplified by a factor of approximately 10 million.\u003C\u002Fp>",{"question":84,"answer":85},"What is Taq polymerase and why is it used in PCR?","\u003Cp>Taq polymerase is a thermostable DNA polymerase originally isolated from Thermus aquaticus, a bacterium that lives in boiling hot springs. Its defining property is heat stability, it remains active at 72°C and survives the 94°C denaturation step without being destroyed. This allows automated PCR cycling without adding fresh enzyme after every cycle. Without a heat-stable polymerase, PCR as an automated process would not be possible.\u003C\u002Fp>",{"question":87,"answer":88},"What is the difference between RT-PCR and real-time PCR?","\u003Cp>These two terms describe different aspects of PCR and are frequently confused. RT-PCR (reverse transcriptase PCR) refers to the template type: it adds a reverse transcription step that converts RNA into complementary DNA before amplification, making it possible to detect RNA viruses such as HIV, hepatitis C, dengue, and SARS-CoV-2. Real-time PCR (quantitative PCR or qPCR) refers to the detection method: fluorescence is measured during each amplification cycle, allowing quantitation of the target. A test can be both simultaneously: the COVID-19 PCR test is technically RT-qPCR, using reverse transcriptase for the RNA template and real-time detection for quantitation.\u003C\u002Fp>",{"question":90,"answer":91},"When should nested PCR be used instead of standard PCR?","\u003Cp>Nested PCR should be used when the target organism is present in very low quantities, below the detection threshold of standard single-round PCR. It uses two successive PCR reactions with two primer sets: outer primers amplify a large fragment first, then inner (nested) primers amplify a smaller specific region within that product. The double amplification dramatically increases sensitivity. Clinical applications include detection of \u003Cem>Rickettsia\u003C\u002Fem> and \u003Cem>Bartonella\u003C\u002Fem> in blood, \u003Cem>M. tuberculosis\u003C\u002Fem> in paucibacillary samples, herpesviruses and enteroviruses in CSF, and \u003Cem>Leishmania\u003C\u002Fem> in tissue.\u003C\u002Fp>",{"question":93,"answer":94},"What are the advantages of PCR over culture in clinical microbiology?","\u003Cp>PCR offers four key advantages over culture. Speed: results in hours rather than days: TB culture takes 6–8 weeks; PCR confirms TB the same day. Sensitivity: detects as few as 1–10 DNA copies per reaction, far below the threshold for culture positivity. Specificity: primers target a defined sequence, identifying the exact organism or resistance gene rather than just confirming growth. Versatility: works on organisms that cannot be cultured (many viruses, some parasites), on degraded specimens (formalin-fixed tissue, dried blood), and on samples with mixed flora where culture is uninterpretable.\u003C\u002Fp>",[66],{"slug":97,"title":98,"description":99,"seoTitle":42,"seoDescription":42,"author":43,"createdDate":100,"lastUpdatedDate":101,"draft":46,"category":47,"image":42,"faq":102,"tags":133},"agarose-gel-electrophoresis","Agarose Gel Electrophoresis: Principle, Procedure, Results","Agarose gel electrophoresis separates DNA fragments from 100 bp to 25 kb by size. Learn the principle, how to cast and run a gel, why supercoiled plasmid runs faster than linear DNA, and how to read a PCR gel against a DNA ladder.","2019-09-13","2026-07-17",[103,106,109,112,115,118,121,124,127,130],{"question":104,"answer":105},"Does agarose gel electrophoresis separate DNA by size or by charge?","By size. Charge determines the direction of travel, because DNA's phosphate backbone is negatively charged and every fragment therefore migrates toward the anode. But DNA has a uniform charge-to-mass ratio, so every fragment experiences the same pull per unit mass, and charge separates nothing. The sieving action of the agarose mesh does all the separating, which is why migration distance reports fragment size.",{"question":107,"answer":108},"Why does DNA move toward the anode?","The phosphate backbone of DNA carries a negative charge at every pH used in the laboratory, making DNA an anion. Anions are attracted to the positive electrode, which is the anode. Unlike proteins, DNA has no isoelectric point to consider, so it always migrates in the same direction.",{"question":110,"answer":111},"What size range can agarose gel electrophoresis resolve?","Roughly 100 base pairs to 25 kilobases, depending on the agarose concentration. Fragments smaller than about 100 bp are better resolved by polyacrylamide gel electrophoresis, and fragments larger than about 25 kb require pulsed-field gel electrophoresis.",{"question":113,"answer":114},"Why does my plasmid preparation show three bands on the gel?","Because a circular plasmid exists in three conformations, and the gel separates by effective size rather than base-pair count. Supercoiled plasmid is tightly wound and compact, so it migrates fastest. Open circular (nicked) plasmid is a relaxed floppy loop with a large effective radius, so it snags in the mesh and migrates slowest. Linear plasmid runs in between. All three contain the same number of base pairs. Three bands from one preparation is normal, not a sign of contamination.",{"question":116,"answer":117},"How do I choose the agarose concentration?","Match the pore size to the fragment size. A low-percentage gel (around 0.5 to 0.8%) has large pores and resolves large fragments, while small fragments run through almost unimpeded. A high-percentage gel (1.5 to 2%) has small pores that resolve small fragments sharply while holding large fragments near the well. Higher percentage does not mean better resolution in general, only better resolution of smaller fragments.",{"question":119,"answer":120},"What is the difference between TAE and TBE buffer?","TAE (Tris-acetate-EDTA) has a lower buffering capacity and will exhaust during long runs, but it resolves large fragments well and the DNA recovered from a TAE gel is clean enough for downstream enzymatic work. TBE (Tris-borate-EDTA) has a much higher buffering capacity and gives sharper resolution of small fragments, but borate inhibits many enzymes and carries over into extracted DNA. Use TAE if you plan to cut the band out and use the DNA, and TBE if you only need to visualize it.",{"question":122,"answer":123},"What is a DNA ladder and why is it needed?","A DNA ladder is a mixture of DNA fragments of known sizes, run in a lane alongside the samples. It converts the height of a band into a number of base pairs. Without a ladder, the position of a band carries no information, because migration distance depends on the gel percentage, the voltage, and the run time.",{"question":125,"answer":126},"My PCR gel shows no band in the patient lane. Is that a negative result?","Only if the positive control produced a band. If the positive control is also blank, the reaction itself failed and the patient's lane carries no information. Likewise, if the negative control shows a band, the run is contaminated and no lane on that gel can be trusted. The controls are always read before the patient's sample.",{"question":128,"answer":129},"What do the dyes in the loading buffer do?","Loading dye serves three purposes. Glycerol makes the sample dense enough to sink to the bottom of the well instead of drifting into the buffer. The colour makes loading easier to see. And the tracking dyes migrate at predictable rates, marking how far the run has progressed. In a 1% agarose gel, bromophenol blue migrates at approximately the position of a 300 to 500 bp fragment and xylene cyanol at approximately 4 kb.",{"question":131,"answer":132},"Is ethidium bromide dangerous, and what can be used instead?","Ethidium bromide intercalates into DNA and is a suspect mutagen and carcinogen, so it requires gloves and regulated disposal. Safer alternatives include SYBR Gold and SYBR Green, which are highly sensitive but expensive, and methylene blue or crystal violet, which are much safer but considerably less sensitive. A separate hazard is the short-wave ultraviolet light used to visualize ethidium bromide, which nicks DNA. If the band is to be excised for cloning, use a long-wave ultraviolet or blue-light transilluminator.",[134],"electrophoresis",{"slug":136,"title":137,"description":138,"seoTitle":42,"seoDescription":42,"author":43,"createdDate":139,"lastUpdatedDate":45,"draft":46,"category":47,"image":42,"faq":140,"tags":156},"multiplex-pcr-principle-applications-and-limitations","Multiplex PCR: Principle, Applications","Multiplex PCR amplifies multiple targets simultaneously in one reaction. Learn primer design considerations, advantages, clinical applications, and limitations in diagnostic microbiology.","2019-12-18",[141,144,147,150,153],{"question":142,"answer":143},"What is multiplex PCR and how does it differ from standard PCR?","Multiplex PCR includes multiple primer pairs in a single PCR reaction, enabling simultaneous amplification of several different target sequences at once. Standard PCR uses one primer pair to detect one target per reaction. Multiplex PCR detects multiple targets — from different organisms or different genes — in the same tube, saving sample volume, reagent cost, and time. Each primer pair produces an amplicon of a specific size, allowing identification of each target by band size on gel or by probe-specific fluorescence in real-time multiplex assays.",{"question":145,"answer":146},"What is the role of the internal amplification control in multiplex PCR?","The internal amplification control is a primer pair directed at a sequence present in all specimens — such as a universal bacterial gene or a human housekeeping gene — included in every multiplex PCR reaction. It serves as a quality gate: if the control amplicon is detected, the PCR conditions were met and a negative result for the test targets can be confidently interpreted as true negative. If the control amplicon is absent, the PCR failed — likely due to inhibitors, degraded nucleic acid, or technical error — and the negative result is uninterpretable. The internal control is what distinguishes a reliable negative from a failed reaction.",{"question":148,"answer":149},"What are the main clinical applications of multiplex PCR in microbiology?","Multiplex PCR is used whenever a clinical syndrome can be caused by multiple pathogens and rapid identification is needed from a limited sample volume. Key applications include: bacterial meningitis panels detecting S. pneumoniae, H. influenzae, and N. meningitidis simultaneously from CSF; respiratory virus panels detecting influenza A, influenza B, RSV, and other respiratory pathogens from nasopharyngeal swabs; gastrointestinal pathogen panels; and sexually transmitted infection panels. The BioFire FilmArray system — which combines nested, multiplex, and singleplex PCR in an automated closed pouch — is the most widely deployed commercial application.",{"question":151,"answer":152},"Why is multiplex PCR challenging to optimise?","Optimising multiplex PCR is challenging because each primer pair has different ideal conditions — melting temperature, MgCl₂ requirement, and amplification efficiency. When multiple primer pairs are combined, more efficient pairs can out-compete less efficient ones, producing strong bands for some targets and weak or absent bands for others (competitive amplification). Primers from different pairs can also interact with each other, forming cross-dimers that consume reagents. Finding annealing temperature and buffer conditions that satisfy all primer pairs simultaneously requires systematic optimisation — adjusting primer concentrations, MgCl₂, and cycling parameters — which becomes increasingly complex as the number of targets increases.",{"question":154,"answer":155},"When is multiplex PCR most useful in clinical practice?","Multiplex PCR is most useful when the differential diagnosis is limited to a defined panel of pathogens causing a specific clinical syndrome. Examples: bacterial meningitis (three main causative organisms), respiratory infections during influenza season (influenza A, B, RSV), and STI panels (gonorrhoea, chlamydia, trichomonas). It is less useful for open-ended differentials where dozens of unrelated pathogens are possible. The key clinical advantage is detecting the causative pathogen from a single limited-volume specimen — particularly important for CSF, vitreous fluid, and neonatal blood where volume constraints prevent running multiple separate reactions.",[66],{"enabled":158,"threads":159,"total":160},true,[],0,[162,168,175,182,188,193,199,204,210,213,220],{"slug":163,"name":43,"description":164,"image":165,"body":166,"postCount":167},"acharya-tankeshwar","Editor-in-chief","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Ftankeshwar-acharya-author-microbeonline.jpg","***Tankeshwar Acharya, MSc (Medical Microbiology)***\n\n*Tankeshwar Acharya is an Assistant Professor in the Department of Microbiology at Patan Academy of Health Sciences (PAHS), Nepal, where he has been teaching and practicing clinical microbiology for over 14 years. He is the founder of Microbe Online, one of the leading free microbiology education resources on the web, covering bacteriology, mycology, parasitology, immunology, and clinical laboratory diagnostics written from direct experience in both the classroom and the diagnostic laboratory.*",477,{"slug":169,"name":170,"description":171,"image":172,"body":173,"postCount":174},"ashma-shrestha","Ashma Shrestha","SEO Copywriter and Science Communicator\nKathmandu, Nepal","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Fashma-shrestha.png","Ashma Shrestha holds a Master of Science in Medical Microbiology from the Institute of Science and Technology (IOST), Tribhuvan University, Nepal, where she developed a strong foundation in virology, molecular biology, and diagnostic microbiology.\n\nShe now works as an SEO Copywriter at Resolution Digital, where she combines her scientific training with research-driven content strategy. She is certified in Google Analytics and Google Business Profile (GBP), and brings a data-informed approach to science communication writing content that is not only accurate but structured to reach and serve the students who need it most.\n\nAt microbeonline, Ashma contributes articles primarily in virology and molecular biology, areas she finds most compelling for their mechanistic depth and their growing clinical relevance. Her writing reflects the same standard the site is built on: factual rigor, clear explanation of the *why* behind microbiology concepts, and content that helps students move from memorization to genuine understanding.\n\nShe is passionate about making complex microbiological concepts accessible without sacrificing accuracy; a skill that sits at the intersection of her scientific training and her professional work in content and SEO.",78,{"slug":176,"name":177,"description":178,"image":179,"body":180,"postCount":181},"sushmita-baniya","Sushmita Baniya","Author \u002F Contributor","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Fsushmita-baniya-1.png","Sushmita Baniya holds an M.Sc. in Medical Microbiology from Tribhuvan University (National College), with a research focus in Genetics and Molecular Biology. She is actively involved in teaching and research in the field of microbiology.",26,{"slug":183,"name":184,"description":178,"image":185,"body":186,"postCount":187},"samikshya-acharya","Samikshya Acharya","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Fsamikshya-acharya.jpeg","Samikshya Sharma completed her postgraduate studies in Medical Microbiology at the Central Department of Microbiology, Tribhuvan University, Nepal. She contributes to Microbeonline with the goal of making foundational and clinical microbiology concepts clear and useful for students in medical, laboratory science, and allied health programs.",20,{"slug":189,"name":190,"description":178,"image":42,"body":191,"postCount":192},"alisha-tripathi","Alisha Tripathi","Alisha Tripathi holds an M.Sc. in Medical Microbiology from National College, Tribhuvan University. With over a year of teaching experience, her academic interests span Molecular Biology, Immunology, and Genetics.",6,{"slug":194,"name":195,"description":196,"image":42,"body":197,"postCount":198},"aastha-shrestha","Aastha Shrestha"," Author \u002F Contributor","Aastha Shrestha is a Biotechnology graduate with an M.Sc. from National College, Tribhuvan University. Her academic interests center on Molecular Biology and Immunology; two fields that are increasingly converging in modern diagnostic and clinical microbiology. \n\nShe contributes to Microbeonline with the goal of making complex concepts in these areas approachable and exam-relevant for students across medical, biotechnology, and laboratory science programs.",9,{"slug":200,"name":201,"description":202,"image":42,"body":42,"postCount":203},"guest-author","Guest Author","Guest Author \u002F Contributor",1,{"slug":205,"name":206,"description":178,"image":207,"body":208,"postCount":209},"srijana-khanal","Srijana Khanal","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Fsrijana-khanal-1.png","Srijana Khanal is a microbiology educator with nearly a decade of teaching experience, including her role as faculty in the Microbiology Department at National College, NIST. \n\nHer time in the classroom has given her a clear sense of where students struggle and what explanations actually work, a perspective that directly shapes how she writes.\n\nHer academic interests span Immunology, Genetics, Basic Sciences, and Research Methodology, and she brings the same rigor to her writing that she brought to teaching. Alongside academic writing, she has a passion for creative writing -- an instinct that shows in her ability to make dense scientific material readable without sacrificing accuracy.\n\nShe contributes to Microbeonline to extend her teaching reach beyond the classroom, helping medical and laboratory science students across the region build a stronger foundation in microbiology.",15,{"slug":211,"name":212,"description":202,"image":42,"body":42,"postCount":203},"dr-poonam-acharya","Dr. Poonam Acharya",{"slug":214,"name":215,"description":216,"image":217,"body":218,"postCount":219},"nisha-rijal","Nisha Rijal","Microbiologist and AMR Specialist Kathmandu, Nepal","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Fnisha-rijal-1.png","Nisha Rijal is a microbiologist with nearly 15 years of frontline diagnostic and surveillance experience at the National Public Health Laboratory (NPHL), national reference laboratory under the Department of Health Services, Nepal. She currently works as an AMR Support Officer at the World Health Organization (WHO), Nepal, where her work focuses on strengthening antimicrobial resistance surveillance systems and translating AMR data into actionable public health response.\n\nHer research, published in peer-reviewed journals and cited over 220 times, spans some of the most clinically significant infectious disease challenges in Nepal and South Asia: antimicrobial resistance trends in *Vibrio cholerae* across an 11-year national surveillance dataset, sero-epidemiology of scrub typhus in patients with acute febrile illness, lower respiratory tract infections in HIV-positive patients, and gonococcal resistance surveillance. She was a contributor to Nepal's National Antimicrobial Resistance Containment Action Plan, a foundational policy document for AMR governance in Nepal. You can find list of [Nisha Rijal's article here in Google Scholar.](https:\u002F\u002Fscholar.google.com\u002Fcitations?user=N-Ruq54AAAAJ&hl=en)\n\nThis depth of experience is visible in her writing at Microbeonline. Her 53 published articles cover bacteriology, parasitology, mycology, immunology, and laboratory techniques, and are consistently among the most detailed and clinically grounded content on the site. She brings to every article the same standard that national reference laboratory work demands: methodological precision, awareness of real diagnostic constraints, and an understanding of what results actually mean for patient care in resource-limited settings.\n\nHer areas of particular expertise include antimicrobial susceptibility testing and resistance mechanism detection, quality assurance in clinical microbiology, and laboratory-based infectious disease surveillance.\n\n---\n\n*Nisha Rijal contributes to Microbeonline in a personal capacity. Her views and writing do not represent the positions of the World Health Organization or any other institution.*",54,{"slug":221,"name":222,"description":223,"image":224,"body":225,"postCount":203},"padma-shrestha","Padma Shrestha","Author","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Fpadma-shrestha.png","Padma Shrestha is from Kathmandu, Nepal. She has completed Masters degree in Medical microbiology from Tribhuvan University. She has great interest in Microbiology and Molecular Biology.",[227,234,240,245,250,255,259,263,267,272,276,281,285,290,295,299,303,307,311,315,319,323,327,332,336,340,344,348,353,358,362,366,370,375,379,383,387,391,395,399,403,407,411,415,419,423,427,431,436,440,444,448,452,456,460,464,468,472,476,480,484,488,492,496,500,504,508,512,515,519,522,525,528,531,534,537,540,543,546],{"slug":228,"name":229,"description":230,"image":231,"body":232,"postCount":233},"gram-negative-cocci","Gram-Negative Cocci and Coccobacilli","Neisseria, Moraxella, Haemophilus and related gram-negative coccal organisms","https:\u002F\u002Fassets.microbeonline.com\u002Ftags\u002Fgram-negative-cocci.png","# Gram Negative Cocci\n\nNeisseria gonorrhoeae, Neisseria meningitides, Moraxella catarrhalis, and other Neisseria spp. are clinically relevant gram-negative cocci.\n\nN. gonorrhoeae is the leading cause of sexually transmitted disease whereas N. meningitides is a leading cause of fatal bacterial meningitis.",14,{"slug":235,"name":236,"description":237,"image":42,"body":238,"postCount":239},"microscopy","Microscopy","Microscope types, components, and microscopy techniques","These are list of blog posts related to microscopy. ",12,{"slug":241,"name":242,"description":243,"image":42,"body":42,"postCount":244},"gram-positive-cocci","Gram-Positive Cocci","Staphylococcus, Streptococcus, Enterococcus, Micrococcus — organisms, diseases, and identification tests",11,{"slug":246,"name":247,"description":248,"image":42,"body":42,"postCount":249},"gram-negative-rods","Gram-Negative Rods (Other than Enterobacteriaceae)","\u003Cp>Gram negative rods other than members of Enterobacteriaceae family such as  Pseudomonas, Acinetobacter and related organisms\u003C\u002Fp>",5,{"slug":251,"name":252,"description":253,"image":42,"body":42,"postCount":254},"gram-positive-rods","Gram-Positive Rods","Bacillus, Clostridium, Listeria, Corynebacterium, Actinomyces and related organisms",8,{"slug":256,"name":257,"description":258,"image":42,"body":42,"postCount":244},"mycobacteria","Mycobacteria","Mycobacterium tuberculosis, leprosy, atypical mycobacteria, and acid-fast organism diagnosis",{"slug":260,"name":261,"description":262,"image":42,"body":42,"postCount":244},"anaerobic-bacteriology","Anaerobic Bacteriology","Anaerobic organisms, anaerobic culture methods, and anaerobic infection diagnosis",{"slug":264,"name":265,"description":266,"image":42,"body":42,"postCount":239},"enterobacteriaceae","Enterobacteriaceae","Identification, differentiation, and clinical significance of Enterobacteriaceae family members",{"slug":268,"name":269,"description":270,"image":42,"body":42,"postCount":271},"spirochetes","Spirochetes","Treponema, Leptospira, Borrelia and spirochetal infections",7,{"slug":273,"name":274,"description":275,"image":42,"body":42,"postCount":209},"food-microbiology","Food Microbiology","Food-borne pathogens, food safety, spoilage, and preservation",{"slug":277,"name":278,"description":279,"image":42,"body":42,"postCount":280},"antimicrobial-susceptibility-testing","Antimicrobial Susceptibility Testing","Methods for testing antibiotic susceptibility in clinical microbiology",21,{"slug":282,"name":283,"description":284,"image":42,"body":42,"postCount":198},"antimicrobials-moa-amr","Antimicrobials (MOA & AMR)","Mechanisms, detection, and clinical significance of antimicrobial resistance",{"slug":286,"name":287,"description":288,"image":42,"body":42,"postCount":289},"sterilization-disinfection","Sterilization and Disinfection","Methods of sterilization and disinfection in healthcare and laboratory settings",10,{"slug":291,"name":292,"description":293,"image":42,"body":42,"postCount":294},"specimen-collection-transport","Specimen Collection and Transport","Collection, handling, and transport of clinical specimens for microbiological testing",27,{"slug":296,"name":297,"description":298,"image":42,"body":42,"postCount":280},"bacterial-structure-physiology","Bacterial Structure and Physiology","Bacterial cell structure, growth, physiology, and environmental factors affecting growth",{"slug":300,"name":301,"description":42,"image":42,"body":302,"postCount":192},"horizontal-gene-transfer","Horizontal Gene Transfer","Articles related to **Horizontal Gene Transfer**",{"slug":304,"name":305,"description":42,"image":42,"body":306,"postCount":289},"chromatography","Chromatography","Information about chromatographic techniques.",{"slug":134,"name":308,"description":309,"image":42,"body":310,"postCount":271},"Electrophoresis","Information about Electrophoresis Techniques ","Detailed information  about Electrophoresis Techniques ",{"slug":66,"name":312,"description":313,"image":42,"body":314,"postCount":192},"PCR Techniques","Information about various types of Polymerase Chain Reaction Techniques ","More detailed information about various types of Polymerase Chain Reaction Techniques ",{"slug":316,"name":317,"description":318,"image":42,"body":42,"postCount":192},"bacteriophage","Bacteriophage","Description about Bacteriophage.",{"slug":320,"name":321,"description":322,"image":42,"body":42,"postCount":192},"malaria","Malaria","It is the collections of articles regarding malarial disease. ",{"slug":324,"name":325,"description":326,"image":42,"body":42,"postCount":192},"anaerobic-culture-techniques","Anaerobic Culture Techniques","Posts related with Anaerobic Culture Techniques.",{"slug":328,"name":329,"description":330,"image":42,"body":42,"postCount":331},"immunoassays","Immunoassays","You will get information about all the diagnostic tests that rely on the specific binding between an antigen and an antibody to detect or quantify a substance.",19,{"slug":333,"name":334,"description":335,"image":42,"body":42,"postCount":271},"biosafety-levels","Biosafety levels ","Articles related to Biosafety Levels",{"slug":337,"name":338,"description":339,"image":42,"body":42,"postCount":249},"environmental-factors","Environmental Factors ","In this case we are talking about growth requirements of microorganisms with deep dive in environmental factors that affect the growth. ",{"slug":341,"name":342,"description":343,"image":42,"body":42,"postCount":192},"pipette","Pipette","Posts related with Pipette. ",{"slug":345,"name":346,"description":347,"image":42,"body":42,"postCount":254},"bacteriology-mcqs","Bacteriology MCQs","This sections lists MCQs in Bacteriology.",{"slug":349,"name":350,"description":351,"image":42,"body":42,"postCount":352},"parasitology-mcqs","Parasitology MCQs","This section lists MCQs in Parasitology.",2,{"slug":354,"name":355,"description":356,"image":42,"body":42,"postCount":357},"virology-mcqs","Virology MCQs","This is the collections of Multiple Choice Questions in Virology.",4,{"slug":359,"name":360,"description":361,"image":42,"body":42,"postCount":249},"mcqs-in-microbiology","MCQs in Microbiology","This section lists the collections of Multiple Choice Questions in General Microbiology Topics. ",{"slug":363,"name":364,"description":365,"image":42,"body":42,"postCount":254},"immunology-mcqs","Immunology MCQs","In this section; we are posting collections of Multiple Choice Questions about Immunology. ",{"slug":367,"name":368,"description":369,"image":42,"body":42,"postCount":198},"microbial-curiosities","Microbial Curiosities","In this clusters, we are posting interesting and unique information about Microorganisms. ",{"slug":371,"name":372,"description":373,"image":42,"body":42,"postCount":374},"bacterial-culture-media","Bacterial Culture Media","Posts related to Bacterial Culture Media. ",22,{"slug":376,"name":377,"description":378,"image":42,"body":42,"postCount":192},"fungal-culture-media","Fungal Culture Media","Posts related to Fungal Culture Media.",{"slug":380,"name":381,"description":382,"image":42,"body":42,"postCount":249},"motility-test","Motility Test","This lists the procedure regarding various tests methods for bacterial motility.",{"slug":384,"name":385,"description":386,"image":42,"body":42,"postCount":289},"bacterial-enumeration","Bacterial enumeration","These posts are related to isolation and enumeration of bacteria. ",{"slug":388,"name":389,"description":390,"image":42,"body":42,"postCount":352},"gram-positive-coccobacillus","Gram-positive coccobacillus","List of Gram Positive Coccobacilli",{"slug":392,"name":393,"description":394,"image":42,"body":42,"postCount":357},"dimorphic-fungi","Dimorphic Fungi","This is about various dimorphic fungi. ",{"slug":396,"name":397,"description":398,"image":42,"body":42,"postCount":271},"bacterial-classification","Bacterial Classification","These posts are related with various approaches used for the classification of Bacteria. ",{"slug":400,"name":401,"description":402,"image":42,"body":42,"postCount":249},"immunofluorescence","Immunofluorescence ","Various Tests related to Immunofluorescence ",{"slug":404,"name":405,"description":406,"image":42,"body":42,"postCount":198},"antibody-mediated-immunity","Antibody-mediated Immunity","This clusters links the articles that are sharing insights about Antibody-mediated immunity. ",{"slug":408,"name":409,"description":410,"image":42,"body":42,"postCount":271},"hypersensitivity","Hypersensitivity","Articles related to Hypersensitivity.",{"slug":412,"name":413,"description":42,"image":42,"body":42,"postCount":414},"haemophilus","Haemophilus",3,{"slug":416,"name":417,"description":418,"image":42,"body":42,"postCount":357},"sexually-transmitted-infections-stis","Sexually transmitted infections (STIs)","This is the clusters of infections that are transmitted sexually. ",{"slug":420,"name":421,"description":422,"image":42,"body":42,"postCount":239},"adaptive-immunity","Adaptive Immunity","Blog posts related to B Cell Immunity and T Cell Immunity.",{"slug":424,"name":425,"description":426,"image":42,"body":42,"postCount":233},"fungal-diagnostics","Fungal Diagnostics","Various methods used for the Diagnosis of Fungal Infections. ",{"slug":428,"name":429,"description":430,"image":42,"body":42,"postCount":249},"laboratory-storage-and-preservation","Laboratory Storage and Preservation","Articles about Laboratory Storage of Antimicrobial Disk, Test organisms and Equipment used for this process. ",{"slug":432,"name":433,"description":434,"image":42,"body":435,"postCount":192},"laboratory-heating-equipment","Laboratory Heating Equipment","A guide to laboratory heating equipment, including hot plates, water baths, Bunsen burners, incubators, and dry baths, and how to choose the right one for each task.","Laboratory heating equipment covers the instruments that warm, melt, incubate, or sterilize samples and media in a microbiology laboratory. Each one delivers heat differently. \n\nA hot plate gives high, direct, dry heat; a water bath gives gentle, even, wet heat up to about 100°C; a Bunsen burner gives an open flame for rapid, very high heat; an incubator holds cultures at a steady temperature over hours or days; and a dry bath heats small tubes without water. Choosing the right one depends on the temperature you need, how precise it must be, and whether the sample can tolerate direct or open-flame heat.\n\nThe articles below cover each piece of heating equipment in detail, including its parts, working principle, uses, and the mistakes that most often go wrong at the bench.",{"slug":437,"name":438,"description":439,"image":42,"body":42,"postCount":198},"laboratory-glassware","Laboratory Glassware","Posts about Laboratory Glassware. ",{"slug":441,"name":442,"description":443,"image":42,"body":42,"postCount":192},"helminths","Helminths","In this section, we are covering properties, life cycle, pathogenesis and laboratory diagnosis of Helminths\u002FHelminthic infestations. ",{"slug":445,"name":446,"description":447,"image":42,"body":42,"postCount":271},"protozoan-parasite","Protozoan Parasite","In this cluster, we are covering protozoan parasites. ",{"slug":449,"name":450,"description":451,"image":42,"body":42,"postCount":203},"tests-for-gram-positive-cocci","Biochemical Tests for Gram Positive Cocci","This is the lists of Biochemical Tests that are used for Gram Positive Cocci. ",{"slug":453,"name":454,"description":455,"image":42,"body":42,"postCount":289},"bacterial-staining-technique","Bacterial Staining Technique","Lists of various staining techniques that are used to stain bacteria. ",{"slug":457,"name":458,"description":459,"image":42,"body":42,"postCount":280},"enzyme-tests","Enzyme Tests","\u003Cp>Various Biochemical Test that are based on enzymatic activity of the microorganisms. \u003C\u002Fp>",{"slug":461,"name":462,"description":463,"image":42,"body":42,"postCount":244},"carbohydrate-utilization","Carbohydrate Utilization","\u003Cp>Various biochemical tests which are related to Carbohydrate fermentation or Utilization\u003C\u002Fp>",{"slug":465,"name":466,"description":467,"image":42,"body":42,"postCount":249},"susceptibility-based-id","Susceptibility-based ID","\u003Cp>These are susceptibility based identification test such as optochin sensitivity, bacitracin sensitivity etc. \u003C\u002Fp>",{"slug":469,"name":470,"description":471,"image":42,"body":42,"postCount":357},"microbial-metabolism","Microbial Metabolism","\u003Cp>Tests about Microbial Metabolism. \u003C\u002Fp>",{"slug":473,"name":474,"description":475,"image":42,"body":42,"postCount":254},"substrate-utilization","Substrate Utilization","\u003Cp>The test in which a non-sugar carbon\u002Fnitrogen source is used or degraded (citrate, malonate, decarboxylases, indole, PAD).\u003C\u002Fp>",{"slug":477,"name":478,"description":479,"image":42,"body":42,"postCount":414},"atypical-pneumonia","Atypical Pneumonia","\u003Cp>Organisms responsible for Atypical Pneumonia. \u003C\u002Fp>",{"slug":481,"name":482,"description":483,"image":42,"body":42,"postCount":249},"antigen","Antigen","\u003Cp>Various articles related to Antigens.\u003C\u002Fp>",{"slug":485,"name":486,"description":487,"image":42,"body":42,"postCount":271},"innate-immunity","Innate Immunity","\u003Cp>Articles related to Innate Immunity. \u003C\u002Fp>",{"slug":489,"name":490,"description":491,"image":42,"body":42,"postCount":357},"respiratory-tract-infection","Respiratory Tract Infection","\u003Cp>In this cluster, you can see various etiological agents that causes respiratory tract infection. \u003C\u002Fp>",{"slug":493,"name":494,"description":495,"image":42,"body":42,"postCount":249},"torch-infection","TORCH Infection","\u003Cp>In this section; you can find articles related with TOCH infection. \u003C\u002Fp>",{"slug":497,"name":498,"description":499,"image":42,"body":42,"postCount":271},"microbiology-for-beginners","Microbiology for Beginners","\u003Cp>These articles are very basic articles, which will share general concepts in Microbiology. \u003C\u002Fp>",{"slug":501,"name":502,"description":503,"image":42,"body":42,"postCount":192},"dna-replication","DNA Replication","\u003Cp>Articles related to DNA and Replication of DNA. \u003C\u002Fp>",{"slug":505,"name":506,"description":507,"image":42,"body":42,"postCount":271},"genetic-code","Genetic Code","\u003Cp>Articles related to Genetic Code.\u003C\u002Fp>",{"slug":509,"name":510,"description":511,"image":42,"body":42,"postCount":249},"molecular-technique","Molecular Technique","\u003Cp>Posts related to Molecular Techniques. \u003C\u002Fp>",{"slug":513,"name":514,"description":42,"image":42,"body":42,"postCount":203},"colorimetric-assay","Colorimetric Assay ",{"slug":516,"name":517,"description":518,"image":42,"body":42,"postCount":249},"pharmaceutical-microbiology","Pharmaceutical Microbiology","\u003Cp>Various articles related to Pharmaceutical Microbiology\u003C\u002Fp>",{"slug":520,"name":521,"description":42,"image":42,"body":42,"postCount":414},"blood-and-immune-cells","Blood and Immune Cells",{"slug":523,"name":524,"description":42,"image":42,"body":42,"postCount":249},"host-pathogen-interaction","Host Pathogen Interaction",{"slug":526,"name":527,"description":42,"image":42,"body":42,"postCount":357},"blood-culture","Blood Culture",{"slug":529,"name":530,"description":42,"image":42,"body":42,"postCount":357},"environmental-microbiology","Environmental microbiology ",{"slug":532,"name":533,"description":42,"image":42,"body":42,"postCount":192},"copromicroscopic-technique","Copromicroscopic Technique",{"slug":535,"name":536,"description":42,"image":42,"body":42,"postCount":414},"quality-control","Quality Control",{"slug":538,"name":539,"description":42,"image":42,"body":42,"postCount":357},"dermatophytes","Dermatophytes",{"slug":541,"name":542,"description":42,"image":42,"body":42,"postCount":414},"viral-hemorrhagic-fevers","Viral Hemorrhagic Fevers",{"slug":544,"name":545,"description":42,"image":42,"body":42,"postCount":357},"h2s-production","H2S Production",{"slug":547,"name":548,"description":42,"image":42,"body":42,"postCount":352},"water-quality-testing","Water Quality Testing"]