Multiplex PCR consists of multiple primer sets within a single PCR mixture to produce amplicons of varying base pairs and sizes that are specific to different DNA sequences. By targeting multiple sequences at once, multiple results are produced in a single test run that otherwise would require several times the reagents and more time to perform.
Advantages of using multiplex PCR include:
- More information with less sample
- Higher throughput
- Cost effective (fewer dNTPs, enzymes, and other consumables)
- Time saving (versus conventional culture methods)
- Less input material required
- More data from limited starting materials
- Increased accuracy of data analysis
- Fewer pipetting errors
- Performed in a closed setting to decrease the possibility of contamination
The possibility of detecting multiple targets in a single sample is particularly important when clinical samples are difficult to collect or are limited in volume (eg, spinal fluid), or when multiple different pathogens can cause the same clinical presentation—making it difficult for clinicians to narrow down the causative pathogen. Thus, multiplex PCR assays are particularly useful for syndromic targets, such as respiratory, gastrointestinal (GI), and the central nervous system. For such targets, there are multiplex PCR assays that are usually offered as a panel which simultaneously detect the pathogens most commonly associated with a particular infectious syndrome using a single specimen.