Key result
The RRCMC assay successfully detected viral pathogens in 44.38% of stool samples from children with acute diarrhea, demonstrating high specificity and a detection limit ranging from 1×10^2 to 1×10^5 copies/ml.
Why the study?
Early and accurate viral identification in children benefits personalized management and reduces serious symptoms, making a high-throughput multiplex real-time RT-PCR method critical to improve routine clinical lab testing.
Cross-Sectional (n=160)
No
The RRCMC method provides a rapid, high-throughput, and specific diagnostic tool for the simultaneous detection of 8 common diarrhea-causing viruses in pediatric patients.
RRCMC may accelerate multiplex virus detection in pediatric labs; leaves open prospective validation before routine use.
Background: Early and accurate identification of infection viruses among children can benefit the personalized medical treatment and management, and reduce the future occurrence of serious symptoms. Thus, it is critical to develop a high-throughput multiplex real-time RT-PCR method to improve the accuracy and efficiency in routine clinical lab tests. Here, we developed a RT-PCR combined with melting curve analysis (RRCMC) method for simultaneous detection of rotavirus A, B, C, norovirus GI and GII, adenovirus, astrovirus and sapovirus. Results: Stool samples were collected from 160 children with acute diarrhea and tested by RRCMC assay. A total of 71 patients were tested positive with norovirus, adenovirus or rotavirus. The RRCMC assay has high specificity. There is no internal cross-reaction through the 8 diarrhea viruses and no cross-reaction of other commonly intestinal pathogens and human genome. The detection limit was ranging from 1×102 to 1×105 copies/ml for each diarrhea virus. Conclusions: In conclusion, the RRCMC method is a suitable rapid clinical test for infection viruses, with the advantages of high-throughput, low cost, high sensitivity, and specificity.
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Li et al. (2020) conducted a cross-sectional in Acute diarrhea (n=160). RT-PCR combined with melting curve analysis (RRCMC) was evaluated on Positive detection rate of 8 diarrhea viruses. The RRCMC assay successfully detected viral pathogens in 44.38% of stool samples from children with acute diarrhea, demonstrating high specificity and a detection limit ranging from 1×10^2 to 1×10^5 copies/ml.
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