Rapid detection of bovine herpesvirus 1 in bovine semen by loop-mediated isothermal amplification (LAMP) assay

被引：13

作者：

Pawar, Sachin S. ^{[1
]}

Meshram, Chetan D. ^{[1
]}

Singh, Niraj K. ^{[2
]}

Sonwane, Arvind A. ^{[1
]}

Saini, Mohini ^{[3
]}

Rautmare, Sunil S. ^{[4
]}

Muglikar, Dushyant M. ^{[5
]}

Mishra, B. P. ^{[1
]}

Gupta, Praveen K. ^{[1
]}

机构：

[1] Indian Vet Res Inst, Div Vet Biotechnol, Izatnagar 243122, Uttar Pradesh, India

[2] Guru Angad Dev Vet & Anim Sci Univ, Sch Anim Biotechnol, Ludhiana 141001, Punjab, India

[3] Indian Vet Res Inst, Ctr Wildlife, Izatnagar 243122, Uttar Pradesh, India

[4] Dept Anim Husb, Dis Invest Sect, Pune 411007, Maharashtra, India

[5] KNP Coll Vet Sci, Dept Microbiol, Shirwal 412801, India

来源：

ARCHIVES OF VIROLOGY | 2014年 / 159卷 / 04期

关键词：

POLYMERASE-CHAIN-REACTION; TIME PCR ASSAY; INFECTED BULLS; VIRUS; BHV-1; DNA; TYPE-1; CATTLE; DIFFERENTIATION; RHINOTRACHEITIS;

D O I：

10.1007/s00705-013-1869-2

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Bovine herpesvirus 1 (BoHV-1) is the most common viral pathogen found in bovine semen, causing numerous reproductive disorders leading to economic losses to the cattle industry. For rapid detection of BoHV-1 in bovine semen, in this study, we applied a loop-mediated isothermal amplification (LAMP) assay. The assay could be completed within 90 min, including total DNA isolation, target amplification, and visual interpretation of positive or negative results with the naked eye. The assay detected as little as 10 fg of BoHV-1 DNA per reaction. The analytical sensitivity of the assay was 0.2 TCID50 BoHV-1 per reaction, which was 100 times more sensitive than conventional PCR and comparable to TaqMan real-time PCR. The applicability of the assay was assessed by analysing 118 semen samples collected from breeding bulls. On comparison with TaqMan real-time PCR, the LAMP assay had a diagnostic sensitivity of 97 %, specificity of 100 %, and accuracy of 99.2 % for detection of BoHV-1 in bovine semen. The LAMP assay developed in this study is a rapid, sensitive, and cost-effective alternative for detection of BoHV-1 in bovine semen.

引用

页码：641 / 648

页数：8

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