Infectious diseases are becoming increasingly complex, with patients often showing overlapping symptoms that make accurate diagnosis difficult. Traditional methods that detect one pathogen at a time are no longer sufficient in today’s fast evolving clinical landscape. There is a growing need for diagnostic tools that can identify multiple pathogens simultaneously, providing rapid and precise results. Modern microbiology technologies, such as multiplex PCR and syndromic testing panels, meet this need by enabling clinicians to detect several infectious agents in a single test, offering faster, more accurate diagnoses and improving patient outcomes.
Multiplex PCR & Syndromic Testing Panels
The multiplex polymerase chain reaction (Multiplex PCR) refers to the use of PCR to amplify several different DNA sequences simultaneously using multiple primers in one assay and one amplification program for all amplicons. By targeting multiple sequences at once, the information is gained from a single assay, reduces the requirement of several tubes and several run times and reagents and provides precise information compared to running multiple separate PCR reactions.
Syndromic testing panels are multiplex molecular tests that use a single patient sample to rapidly identify multiple potential pathogens and resistance genes simultaneously. They are designed to diagnose infections with overlapping symptoms, like respiratory or gastrointestinal illnesses, by testing for many different bacteria, viruses, fungi, and parasites at once. This approach reduces the time to diagnosis, and helps healthcare providers make faster, more appropriate treatment decisions compared to traditional, single-organism testing. Syndromic diagnostic panels are now commercially available to aid in the diagnosis of common, serious infections that affect the bloodstream, respiratory, gastrointestinal, and central nervous systems. (1)
Clinical Evaluation & Impact on Patient Care:
In a recent clinical evaluation, the performance of four multiplex real-time PCR panels designed to detect infectious agents directly from blood, cerebrospinal fluid (CSF), respiratory, and stool samples were analysed. This panel was tested on infected sample of 1,929 patients and results were compared with traditional laboratory methods such as culture and standard PCR and in result a high diagnostic accuracy demonstrated with multiplex PCR Panels, confirming the efficiency of the multiplex PCR. (2)
- 97.3% sensitivity and 96.3% specificity for respiratory pathogens.
- 94.3% sensitivity and 97.9% specificity for gastrointestinal pathogens.
- 96.4% sensitivity and 96.8% specificity for CNS infection detection.
- 97.1% sensitivity and 99.3% specificity for pathogens in positive blood cultures.
Multiplex PCR was able to produce results in approximately 3 hours as compared to 24–72 hours for culture-based diagnosis. This method allows clinicians to start targeted treatment earlier which reduces antibiotic usage and improve infection control decisions.
Case study: Rapid diagnosis of sepsis with TaqMan-based multiplex real-time PCR
Early identification is essential for survival in cases of sepsis, a potentially fatal bloodstream infection. Effective therapy is frequently delayed by two to three days to identify the virus using traditional blood culture methods. On the other hand, this study’s TaqMan-Based Multiplex Real-Time PCR assay identify bacterial DNA straight from blood in a matter of hours. The test showed great sensitivity by identifying infections in 18 out of 30 suspected sepsis patients by focusing on the conserved 16S rDNA region present in most bacteria, whereas only 7 were recognized by blood culture. This demonstrates that Multiplex PCR is a useful technique for prompt and precise sepsis diagnosis since it can detect twice as many infections and much more quickly (3).
Abdos Innovation in Action:
Integrating Multiplex PCR requires high accuracy and contamination-free sample handling, as even a minor carryover of nucleic acids can lead to false positives or incorrect pathogen identification, and there to ensure reliable results, Abdos filter pipette tips play a critical role in maintaining aseptic liquid handling throughout the workflow. Abdos Filter Tips are manufactured using 100% pure porous polyethylene (PE) super filters that create a physical barrier to prevent the passage of aerosols and liquid droplets. This design protects both the sample and the pipette from cross-contamination, making them essential for sensitive workflows such as PCR, Multiplex PCR, qPCR, RT-PCR, NGS, and viral diagnostics.
To conclude Multiplex PCR and syndromic diagnostic panels are the major breakthroughs to test infectious disease by providing rapid, accurate, and comprehensive pathogen detection. Utilization of these panel have accelerated the clinical decision-making, enabling targeted therapy, reducing exposure of unnecessary antibiotic and improve patient outcomes especially in time-sensitive conditions. In addition, to exhibit these methods, a strict contamination control is needed, placing high quality filter pipette tips as an essential part of every molecular diagnostics workflow.
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