A rapid, simple and sensitive loop-mediated isothermal amplification method to detect Anaplasma bovis in sheep and goats samples

Parasitol Int. 2018 Feb;67(1):70-73. doi: 10.1016/j.parint.2017.03.005. Epub 2017 Mar 27.

Abstract

A loop-mediated isothermal amplification (LAMP) technique has been widely used in detecting the nucleic acid of various pathogenic bacteria. In this study, a set of four LAMP primers was designed to specifically test Anaplasma bovis. The LAMP assay was performed at 62°C for 60min in a water bath. The specificity was confirmed by amplifying A. bovis isolate, while no cross reaction was observed with other five pathogens (Anaplasma bovis, Anaplasma phagocytophilum, Theileria luwenshuni, Babesia motasi and Schistosoma japonicum). The sensitivity of LAMP was 5×100copies/μL, 100 times more than that of conventional PCR (5×102copies/μL). Of 120 blood DNA extracted from sheep and goats field samples, 81 (67.5%), 22 (18.3%) and 43 (35.8%) were positively detected by LAMP, conventional PCR and nested PCR, respectively. The findings indicated that the developed LAMP assay is a new convenient tool for rapid and cost-effective detection of A. bovis.

Keywords: 16S rRNA; Anaplasma bovis; LAMP assay.

MeSH terms

  • Anaplasma / isolation & purification*
  • Anaplasmosis / diagnosis*
  • Anaplasmosis / parasitology
  • Animals
  • DNA, Protozoan / genetics
  • Goat Diseases / diagnosis*
  • Goat Diseases / parasitology
  • Goats
  • Nucleic Acid Amplification Techniques / economics
  • Nucleic Acid Amplification Techniques / veterinary*
  • Polymerase Chain Reaction / veterinary
  • RNA, Ribosomal, 16S / genetics
  • Sensitivity and Specificity
  • Sheep
  • Sheep Diseases / diagnosis*
  • Sheep Diseases / parasitology

Substances

  • DNA, Protozoan
  • RNA, Ribosomal, 16S