ArticleBMC infectious diseases2023
Development and clinical validation of a one-step pentaplex real-time reverse transcription PCR assay for detection of hepatitis virus B, C, E, Treponema pallidum, and a human housekeeping gene.
Article in BMC infectious diseases, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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7 citing papers in PubMed, 6 citations in OpenAlex.
- Sensitive and Rapid Dual One-Tube Recombinase Aided PCR Assays for the Simultaneous Detection of Cytomegalovirus, Epstein-Barr Virus, Human Herpesvirus 6, and Herpes Simplex Virus 1/2 in Whole Blood From Children Undergoing Hematopoietic Stem Cell Transplantation.Journal of medical virology · 2026Article
- Article
- Viral hepatitis as a differential diagnosis of yellow fever suspected cases in Cameroon: Prevalence and molecular characterization.IJID regions · 2025Article
- A Perspective on Wastewater and Environmental Surveillance as a Public Health Tool for Low- and Middle-Income Countries.Microorganisms · 2025Article
- Current knowledge on the epidemiology and detection methods of hepatitis E virus in China.Virology journal · 2024Review
- Spotted fever diagnosis using molecular methods.Revista da Sociedade Brasileira de Medicina Tropical · 2024Review
- Rapid and sensitive detection of methicillin-resistantFrontiers in microbiology · 2024Article
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9 authors at 4 institutions in 1 country.
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Abstract
backgroundWith the safety of blood transfusion being a major public health concern, the development of a rapid, sensitive, specific, and cost-effective multiplex PCR assay for simultaneous detection of hepatitis B virus(HBV), hepatitis C virus (HCV), hepatitis E virus (HEV), and Treponema pallidum(T. pallidum) in blood is crucial.
methodsFive primer pairs and probes were designed towards conserved regions of target genes and used to establish a one-step pentaplex real-time reverse transcription PCR(qRT-PCR) assay for simultaneous detection of HBV, HCV, HEV, T. pallidum, and RNase P(housekeeping gene), providing sample quality check. The clinical performance of the assay was further determined with 2400 blood samples from blood donors and patients in Zhejiang province, and compared the results with commercial singleplex qPCR and serological assays.
resultsThe 95% limit of detection(LOD) of HBV, HCV, HEV, and T. pallidum were 7.11 copies/µL, 7.65 copies/µL, 8.45 copies/µL, and 9.06 copies/µL, respectively. Moreover, the assay has good specificity and precision. Compared to the singleplex qPCR assay, the novel assay for detecting HBV, HCV, HEV, and T. pallidum presented 100% clinical sensitivity, specificity, and consistency. Several discrepant results between serological and pentaplex qRT-PCR assays were found. Of 2400 blood samples, there were 2(0.08%) HBsAg positive samples, 3(0.13%) anti-HCV positive samples, 29(1.21%) IgM anti-HEV positive samples and 6(0.25%) anti-T. pallidum positive samples proven negative in nucleic acid detection. 1(0.04%) HBV DNA positive sample and 1(0.04%) HEV RNA positive sample were detected negative by serological testing.
conclusionsThe developed pentaplex qRT-PCR is the first assay on simultaneous, sensitive, specific, and reproducible detection of HBV, HCV, HEV, T. pallidum, and RNase P in a single tube. It could detect pathogens in blood during the window period of infection and is a good tool for effectively screening blood donors and early clinical diagnosis.
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