Probe-free and sensitive detection of diarrhea-causing pathogens using RT-PCR combined high resolution melting analysis.

Autor: Wang HB; State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China. Electronic address: wanghb1013@hotmail.com., Mo QH; State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China., Wang Q; State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China., Wu BM; State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China., Feng ZL; State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China., Lin JC; State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China., Yang Z; State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China.
Jazyk: angličtina
Zdroj: Biologicals : journal of the International Association of Biological Standardization [Biologicals] 2016 Sep; Vol. 44 (5), pp. 360-6. Date of Electronic Publication: 2016 Jul 25.
DOI: 10.1016/j.biologicals.2016.06.012
Abstrakt: Rapid and sensitive diagnostic methods are needed to help physicians make faster and better treatment decision for patients suffered from diarrhea. In the present study, a probe-free and sensitive RT-PCR combined high resolution melting analysis (HRMA) assay was established successfully for the detection of four major diarrhea-causing pathogens. The lower limit of detection of the assay were 10(0), 10(2), 10(0) and 10(3) copies/reaction for rotaviruses group A, astroviruses serotype 1, noroviruses genogroup II, and sapoviruses genegroup I, respectively, which were 1000-fold, 10-fold, 1000-fold and 10-fold more sensitive than conventional RT-PCR assay developed in parallel and comparable to or higher than commercially available real-time RT-PCR assay. Blinded sample evaluation showed that the assay was 100% concordant to both conventional RT-PCR and commercial real-time RT-PCR, indicating high reliability of the new assay. Therefore, the assay could provide a valuable platform for the probe-free and sensitive diagnosis of these pathogens.
(Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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