Multiplex PCR consists of multiple primer sets within a single PCR mixture to produce amplicons of varying base pairs and sizes specific to different DNA sequences. Targeting various sequences at once produces multiple results 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 significant when clinical samples are complex to collect or are limited in volume (e.g., spinal fluid) or when numerous 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, multiplex PCR assays are usually offered as a panel that simultaneously detects the pathogens most commonly associated with a particular infectious syndrome using a single specimen.
