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Open Access Methodology

Development and evaluation of a loop-mediated isothermal amplification assay for the rapid detection of porcine cytomegalovirus under field conditions

Jin-Long Yang1, Su-Hui Zhang1, Zuo-Hua Liu1, Rui Yang1, Yong Huang1 and Ming Wen2*

Author Affiliations

1 Chongqing Academy of Animal Science, Chongqing, 402460, China

2 Animal Science College of Guizhou University, Guiyang, Guizhou, 550025, China

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Virology Journal 2012, 9:321  doi:10.1186/1743-422X-9-321

Published: 29 December 2012

Abstract

Background

Porcine cytomegalovirus (PCMV) induces silent infection in adult pigs but more frequently causes fatal, generalized infection in newborn piglets. This study aimed to develop a new loop-mediated isothermal amplification (LAMP) method for the sensitive, rapid, and inexpensive detection of PCMV under field conditions.

Methods

Tissue obtained from nine-week-old PCMV-free Landrace pigs or pig samples from postmortem examinations were analyzed. The samples were found to have clinical signs and lesions consistent with inclusion body rhinitis. Six specific primers were designed by targeting the PCMV DNA polymerase (DPOL) DNA. The LAMP reaction was optimized in a water bath. The sensitivity and specificity of LAMP and polymerase chain reaction (PCR) were compared.

Results

PCMV DNA was amplified at 65°C, and the result could be detected as early as 30 min into the reaction. Positive reactions could be visualized by the naked eye as a color change brought on by the addition of SYBR Green. The sensitivity and specificity of LAMP were found to be similar to those of the PCR.

Conclusions

LAMP is a high-throughput technique for the detection of PCMV and has a high specificity, sensitivity and simplicity; these factors make it suitable for detection of PCMV under field conditions.

Keywords:
Porcine cytomegalovirus; Loop-mediated isothermal amplification; Landrace pigs; Field detection