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In order to establish a rapid detection method for  Mycoplasma ovipneumoniae , this study used the loop-mediated isothermal amplification (LAMP) technique to carry out nucleic acid amplification and chromatographic visualization via a lateral flow dipstick (LFD) assay. The  M. ovipneumoniae  elongation factor  TU  gene ( EF-TU ) was detected using a set of specific primers designed for the  EF-TU  gene, and the  EF-TU  FIP was detected by biotin labeling, which was used in the LAMP amplification reaction. The digoxin-labeled probe specifically hybridized with LAMP products, which were visually detected by LFD. Here, we established the  M. ovipneumoniae  LAMP-LFD rapid detection method and tested the specificity, sensitivity, and clinical application of this method. Results showed that the optimized LAMP performed at 60 °C for 60 min, and LFD can specifically and visually detect  M. ovipneumoniae  with a minimum detectable concentration at 1.0 × 10 2  CFU/mL. The sensitivity of LAMP-LFD was 1000 times that of the conventional PCR detection methods, and the clinical lung tissue detection rate was 86% of 50 suspected sheep infected with  M. ovipneumoniae . In conclusion, LAMP-LFD was established in this study to detect  M. ovipneumoniae , a method that was highly specific, sensitive, and easy to operate, and provides a new method for the prevention and diagnosis of  M. ovipneumoniae  infection.   Electronic supplementary material  The online version of this article (10.1007/s11274-019-2601-5) contains supplementary material, which is available to authorized users.

world journal of microbiology and biotechnology incorporating the mircen journal of applied microbiology and biotechnology/world journal of microbiology and biotechnology

Loop-mediated isothermal amplification-lateral-flow dipstick (LAMP-LFD) to detect Mycoplasma ovipneumoniae