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Comparison of three molecular detection methods for detection of Trichinella in infected pigs

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Abstract

Different molecular detection methods require diverse molecular platforms, but there is no uniform standard for people to reference in the detection of Trichinella. In this study, real-time PCR, loop-mediated isothermal amplification (LAMP), and conventional PCR were developed for the detection of Trichinella by targeting mitochondrial large subunit ribosomal DNA (mt-lsrDNA). We compared the performance of the three newly developed assays. The results revealed that the detection limits of the real-time PCR, LAMP, and conventional PCR assays were 10 and 100 fg/μL and 1 pg/μL of Trichinella spiralis genomic DNA, respectively. The assays were used in the detection of Trichinella in the field. A total of 192 samples were obtained from pigs: 75 samples from free range farming and 117 from intensive feeding factory. The infection rate was 8/192 (4.2 %), 7/192 (3.6 %), and 1/192 (1.0 %) through the real-time PCR, LAMP, and conventional PCR assays, respectively. These data indicate that Taqman real-time PCR was a rapid, specific, and sensitive tool as a preferred option for investigation of valuable samples, but that LAMP assay was closed tube, highly sensitive, cost-effective, rapid, easy-to-perform, and was the optimal choice for detection of Trichinella in the field. The results of a model of experimental infection in mice indicated that spleen can be used as sampling site for the detection of early T. spiralis infection. However, the diaphragm and myocardium were the most suitable sampling sites for the detection of T. spiralis.

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Acknowledgments

This work was supported by the Special Fund for Quarantine Research in the Public Interest (Grant No. 201110039).

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Authors and Affiliations

  1. Key Laboratory of Animal Parasitology of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China

    Zhibing Lin, Jie Cao, Houshuang Zhang, Yongzhi Zhou & Jinlin Zhou

  2. South China Agricultural University, College of Veterinary Medicine, Guangzhou, 510642, China

    Zhibing Lin & Guoqing Li

  3. Shandong Entry-Exit Inspection and Quarantine Bureau, Qingdao, 266002, China

    Mingjun Deng

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  1. Zhibing Lin
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  2. Jie Cao
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  3. Houshuang Zhang
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  4. Yongzhi Zhou
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  6. Guoqing Li
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  7. Jinlin Zhou
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Correspondence to Guoqing Li or Jinlin Zhou.

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Lin, Z., Cao, J., Zhang, H. et al. Comparison of three molecular detection methods for detection of Trichinella in infected pigs. Parasitol Res 112, 2087–2093 (2013). https://doi.org/10.1007/s00436-012-3267-1

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