African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12392

Full Length Research Paper

Rapid single-tube splice variants typing of the BF gene based on dual-primer RT-PCR amplification that influence resistance/susceptibility to Marek’s disease in chicken

Yuan-chang Jin
  • Yuan-chang Jin
  • College of Biology and Agriculture (College of Food Science and Technology), Zunyi Normal College, Zunyi 563006, China.
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Yu-feng Li
  • Yu-feng Li
  • College of Agriculture and Food Engineering, Baise University, Baise 533000, China.
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Liang Huang
  • Liang Huang
  • Poultry Science and Health, Guangxi University, Nanning 530004, China.
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Jia-jun Zhou
  • Jia-jun Zhou
  • College of Biology and Agriculture (College of Food Science and Technology), Zunyi Normal College, Zunyi 563006, China.
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Xue-fang Zhang
  • Xue-fang Zhang
  • College of Biology and Agriculture (College of Food Science and Technology), Zunyi Normal College, Zunyi 563006, China.
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Ran Ma
  • Ran Ma
  • College of Biology and Agriculture (College of Food Science and Technology), Zunyi Normal College, Zunyi 563006, China.
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Meng-lin Lu
  • Meng-lin Lu
  • College of Biology and Agriculture (College of Food Science and Technology), Zunyi Normal College, Zunyi 563006, China.
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Mei-lin Hao
  • Mei-lin Hao
  • College of Biology and Agriculture (College of Food Science and Technology), Zunyi Normal College, Zunyi 563006, China.
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Gang Zeng
  • Gang Zeng
  • College of Biology and Agriculture (College of Food Science and Technology), Zunyi Normal College, Zunyi 563006, China.
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Bo-ping Zeng
  • Bo-ping Zeng
  • College of Biology and Agriculture (College of Food Science and Technology), Zunyi Normal College, Zunyi 563006, China.
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  •  Received: 24 October 2021
  •  Accepted: 07 January 2022
  •  Published: 31 January 2022

Abstract

A new effective splice variants typing based on multiplex allele-specific dual-primer RT-PCR assay was developed in a single tube for the rapid detection of the exon 7 splice variant of the BF gene. With 2 pairs of primers, one pair was used for amplifying cDNA fragments containing exon 7 of the BF gene, the other does not contain exon 7 of the BF gene. The templates were amplified in one tube and the type of splice variants was determined by the length of products to be extended and by analysis of nucleotide sequences of these BFs. Results obtained for all samples showed 100% accuracy compared to those obtained with a semi-nested PCR (snPCR) assay of 100% accuracy, but which need two round PCR assay. The dual-primer RT-PCR assay was more rapid and easy to operate than the snPCR assay.

 

Key words: BF gene (chicken MHC class I gene), Marek’s disease (MD), splice variants, resistance, chickens.