Development of Loop-mediated Amplification for Detection of Eimeria stiedai

doi:10.3969/j.issn.1674-5817.2018.03.003

Abstract

Abstract: Objective Loop-mediated amplification (LAMP) approaches was developed for detecting Eimeria stiedai (E.stiedai).Methods Specific primers were designed and synthesized according to a part of the sequence of ITS1-5.8sRNA-ITS2 region of E.stiedai.The LAMP methods of detecting E.stiedai was established by optimizing the reaction conditions.Results In 25 µL reaction system,the optimal dosage of Bst polymerase is 1.5 µL,and the optimal dosage of DNA is 75 ng.The optimal concentration of ring primers,internal primers and external primers is 0.8 µmol/L,1.6 µmol/L and 0.15 µmol/L respectively.The target E.stiedai strip can be specifically amplified when reacted for 1h at 63 ℃.The sensitivity of LAMP was high for determing E.stiedai,and this method could specifically detect the DNA of E.stiedai. Conclusions The LAMP method for detecting E.stiedai has been established,which provides a new method for rapid detection of E.stiedai infection.

Key words: E. stiedai, PCR, Loop-mediated amplification (LAMP), Sensitivity, Specificity

CLC Number:

R-332Q95-33

WEN Fu-li. Development of Loop-mediated Amplification for Detection of Eimeria stiedai[J]. Laboratory Animal and Comparative Medicine, 2018, 38(3): 176-181.

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