Multiplex Pcr Assay
Multiplex Pcr Assay - Related Videos
10:15
Multiplex PCR and Reverse Line Blot Hybridization Assay (mPCR/RLB)
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Cited by 21 •
2011
An inexpensive, high throughput method for simultaneous detection of up to 43 molecular targets is described. Applications of mPCR/RLB include microbial typing and detection of multiple pathogens from clinical samples.
09:03
Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
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Cited by 18 •
2013
We demonstrate a simple multiplex PCR assay for quick-screening and typing of Staphylococcal Cassette Chromosome mec (SCCmec) types I-V for methicillin-resistant Staphylococcus aureus, and provide some of the vital steps and procedural nuances that make it successful for adapting this assay to individual laboratories.
03:41
Agarose Gel Electrophoresis of DNA Amplicons Post PCR: A Method to Analyze Products of Multiplex PCR and Evaluate PCR Reaction Success
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2025
In this video, we demonstrate the separation of bacterial PCR-amplified DNA using agarose gel electrophoresis. Agarose gel functions as a molecular sieve, enabling the negatively-charged DNA to migrate based upon their size under an applied electric field.
07:27
High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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Cited by 13 •
2011
Here we describe an optimized multiplex reverse transcriptase quantitative PCR (qRT-PCR) protocol in combination with a microfluidic platform as a cost and time effective high-throughput screening tool for microRNA (miRNA) expression levels, especially when working with limited amounts of sample.
05:22
A Multiplex Serological Assay for the Detection of Antibody Responses to Arboviruses
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2025
Multiplex fluorescence immunoassays are becoming a reference for serology testing. Their application, however, is limited by the complexity and time-consuming bead-analyte coupling processes. This report presents a semi-automated, reproducible multiplex microsphere bead coupling protocol that is comparable to the standard method while considerably reducing time to results.
FAQs
A multiplex PCR assay relies on each primer pair recognizing its complementary nucleic-acid target during the annealing stage. Subsequent extension copies only the targets matched by those primers, while repeated denaturation, annealing, and extension cycles increase the amount of each product. This coordinated selectivity allows several targets to be examined in one reaction rather than processed separately.
Distinct amplicon sizes or fluorescent probes provide the separation needed to tell products apart. Size-based distinction relies on amplified fragments having different lengths, whereas probe-based distinction uses fluorescent probes to identify products. This readout step is essential because amplification produces multiple products; discrimination connects each observed fragment or signal to its corresponding pathogen or genetic marker.
Combining targets in one reaction conserves sample and reagents while reducing processing time. It also increases laboratory throughput, which is valuable when investigators need to examine several pathogens or genetic markers from limited material. The practical benefit is not simply more targets, but a more efficient investigation that can compare multiple relevant signals within the same specimen.
A typical workflow brings the sample and selected primer pairs into a single reaction, then applies repeated denaturation, primer annealing, and extension cycles. After amplification, investigators distinguish the resulting products by their sizes or by fluorescent probes. This sequence links target selection, controlled copying, and product identification, allowing several pathogen or marker results to be generated from one reaction.
It is particularly useful when a specimen may contain more than one pathogen or when rapid identification must cover several possible infectious agents. The assay supports co-infection assessment by examining multiple targets together, while its efficient format helps laboratories process investigations more quickly. These capabilities make it relevant to studies requiring broad pathogen detection while conserving sample material.
Multiplex PCR can include microbial genetic targets used for antimicrobial-resistance screening alongside targets used for pathogen identification. This pairing allows an investigation to examine the infectious agent and relevant resistance markers within the same molecular testing strategy. The approach strengthens laboratory characterization of infection while preserving the assay’s broader ability to assess several genetic features together.
The assay can be configured to analyze host genes or microbial genes, extending an investigation beyond identification of an infectious organism. In immunology and infection research, these targets allow molecular information from the host or microbe to be examined within the same broader laboratory strategy. Multiplexing also conserves sample, reagents, and processing time during such investigations.