Establishment of TaqMan Real-time PCR Method for Detection of Paranosema locustae

doi:10.16409/j.cnki.2095-039x.2017.06.012

Abstract

Abstract: Paranosema locustae, an entomopathogen of locusts, is widely used in biocontrol of locusts. The small subunit ribosomal RNA gene of P. locustae was used as the target molecule for designing specific primers and TaqMan probe. pMD18-T, the constructed recombinant plasmid was used to prepare standard agent for fluorescent quantitative PCR amplification. Through optimization of PCR reaction system, linear range of the detection method was determined, and sensitivity and repeatability of the method were evaluated. Finally, a fluorescent quantitative PCR detection method was established successfully. The detectable sensitivity of this method reached 1.29×10⁵ copies/L. Three repeated detection trials showed that the variation coefficients within batches and between batches were both less than 1%. These results indicated that the established fluorescent quantitative PCR based on TaqMan probe technology is an accurate, sensitive and easy to operate method for P. locustae detection.

Key words: Paranosema locustae, fluorescent quantitative PCR, TaqMan probe

CLC Number:

S476.1

HU Hongxia, ZHENG Qiuying, YE Xiaofang, ZHAO Bei, JI Rong. Establishment of TaqMan Real-time PCR Method for Detection of Paranosema locustae[J]. journal1, 2017, 33(6): 796-802.

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