SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses

被引：82

作者：

Escutenaire, S.

Mohamed, N.

Isaksson, M.

Thoren, P.

Klingeborn, B.

Belak, S.

Berg, M.

Blomberg, J.

机构：

[1] Natl Vet Inst, Dept Virol, S-75189 Uppsala, Sweden

[2] Uppsala Univ, Acad Hosp, Dept Med Sci, Sect Clin Virol, Uppsala, Sweden

[3] Swedish Univ Agr Sci, Dept Biomed Sci & Vet Publ Hlth, Sect Parasitol & Virol, Uppsala, Sweden

来源：

ARCHIVES OF VIROLOGY | 2007年 / 152卷 / 01期

关键词：

D O I：

10.1007/s00705-006-0840-x

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Coronaviruses are etiologic agents of respiratory and enteric diseases in humans and in animals. In this study, a one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on SYBR Green chemistry and degenerate primers was developed for the generic detection of coronaviruses. The primers, designed in the open reading frame 1b, enabled the detection of 32 animal coronaviruses including strains of canine coronavirus, feline coronavirus, transmissible gastroenteritis virus (TGEV), bovine coronavirus (BCoV), murine hepatitis virus (MHV) and infectious bronchitis virus (IBV). A specific amplification was also observed with the human coronaviruses (HCoV) HCoV-NL63, HCoV-OC43, HCoV-229E and severe acute respiratory syndrome coronavirus (SARS-CoV). The real-time RT-PCR detected down to 10 cRNA copies from TGEV, BCoV, SARS-CoV and IBV. In addition, the assay exhibited a high sensitivity and specificity on clinical samples from different animal species. The developed assay represents a potential tool for laboratory diagnostics and for detecting still uncharacterized coronaviruses.

引用

页码：41 / 58

页数：18

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