Isothermal cross-priming amplification implementation study

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Cross-priming amplification (CPA) was evaluated for the early detection of norovirus (NV), rotavirus A (RV-A), enteric adenovirus (EAdV) and astrovirus (AstV). The analytical sensitivity of the CPA assay was 103 copies ml−1 for NV, RV-A and AstV detection and 104 copies ml−1 for EAdV detection. For each of the four pathogens, the positive detection rate by CPA was similar to real-time PCR methods and higher than the rate observed in an ELISA. The detection coincidence rates of CPA and RT-PCR for NV, RV-A, EAdV and AstV were 98, 99, 99 and 100%, respectively. All CPA assays were negative in 89 healthy control samples. These results demonstrate the high analytical sensitivity and specificity of the CPA assay. CPA assays are relatively straightforward to perform, and such assays represent a potential detection method for locations in which resources are limited.Significance and Impact of the StudyOver one billion people suffer from diarrhoeal diseases every year. This leads to significant morbidity and mortality, particularly the children under five. Rapid and specific detection of the pathogens that cause diarrhoeal diseases would be advantageous, enabling rapid treatment and management of the spread of pathogens. Here, a fast, cross-contamination-proof and user-friendly nucleic acid isothermal amplification method called cross-priming amplification (CPA) was used to test four pathogens with high analytical sensitivity and specificity. The results indicate that CPA has great potential for improving medical diagnostics, particularly in resource-limited areas.

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