title: Probe-free and sensitive detection of diarrhea-causing pathogens using RT-PCR combined high resolution melting analysis.
abstract: 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.
authors.affiliation: 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.
authors.forename: Hai-Bo
authors.initials: HB
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China.
authors.forename: Qiu-Hua
authors.initials: QH
authors.lastname: Mo
authors.affiliation: State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China.
authors.forename: Bi-Mei
authors.initials: BM
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China.
authors.forename: Zi-Li
authors.initials: ZL
authors.lastname: Feng
authors.affiliation: State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China.
authors.forename: Ji-Can
authors.initials: JC
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, Guangdong, China.
authors.forename: Ze
authors.initials: Z
authors.lastname: Yang
mesh_terms: D003967:Diarrhea / Q000175:diagnosis / Q000235:genetics / Q000821:virology; D005260:Female; D006801:Humans; D008297:Male; D009691:Nucleic Acid Denaturation; D012327:RNA Virus Infections / Q000175:diagnosis / Q000235:genetics; D012328:RNA Viruses / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D012367:RNA, Viral
keywords: Diagnosis; Diarrhea; High resolution melting analysis; RT-PCR