Sensitive and rapid detection of Mycoplasma capricolum subsp. capripneumoniae by Loop-mediated isothermal amplification

Abstract

A set of four specific primers was designed by targeting the H2 gene sequences of Mycoplasma capricolum subsp. capripneumoniae (MCCP). Using Bst DNA polymerase, the products were amplified for 60 min at 65°C in a simple water bath. Compared with a polymerase chain reaction (PCR) test that targets the H2 gene sequences of MCCP, the sensitivity of the loop-mediated isothermal amplification (LAMP) assay was higher (approximately 0.75 fg DNA per reaction). The LAMP products could be visualized by agar gel electrophoresis. There were no cross reactions with other strains in the Mycoplasma mycoides cluster, which indicates the high specificity of the LAMP procedure. The LAMP assay was able to detect MCCP in tissue. Key words: Mycoplasma capricolum subsp. Capripneumoniae, loop-mediated isothermal amplification, rapid detection.