Detection of Porcine Parvovirus using Loop-mediated Isothermal Amplification

ISSN: 0973-7510

E-ISSN: 2581-690X

Fang Liu¹ , JinLei Li¹, HongYing Chen², HongYing Zhang², XueBing Wang², BaoAn Cui and ZhanYong Wei²

¹The College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan – 450 002, P.R.China.

²Henan Key Laboratory for Animal-derived Food Safety, Henan Agricultural University of Henan Province, Zhengzhou, Henan – 450 002, P.R.China.

J Pure Appl Microbiol. 2013;7(1):311-315

Received: 11/02/2013 | Accepted: 02/03/2013 | Published: 31/03/2013

Abstract

Loop-mediated isothermal amplification (LAMP) is a novel, sensitive and rapid technique for detection of genomic DNA or RNA. In this report, the LAMP assay reported can provide a more sensitivity and specificity of PPV test, comparing with PCR assay. The results showed that the amplification could be finished in 1 h under isothermal condition at 63! by employing a set of four primers targeting NS1 gene of PPV. The end-product of the technique is a white precipitate of magnesium pyrophosphate that is visible without the use of gel electrophoresis. The detection limit of the LAMP method was as low as 5 copies and was 25 times more sensitive than that of PCR assay. When detecting samples infected with PPV, PRV, PRRSV, PCV2, JEV, HCV by LAMP assay, only sample infected with PPV was amplified. The detection rates of PCR and LAMP assay were 75% and 100% for the 25 clinical samples, respectively. The LAMP assay is an easy, rapid and sensitive method and has great potential for use in the detection of PPV in the laboratory and the farm.

Keywords

Loop-mediated isothermal amplification (LAMP) assay, Porcine parvovirus, Sensitive and rapid detection

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