Sensitivity comparison of PCR primers for detecting Wolbachia in spider mites

Abstract

Some primers occasionally fail to amplify Wolbachia DNA by a standard PCR from spider mites. In cross experiments, it is often necessary to detect Wolbachia from single individuals. Thus, six pairs of PCR primers were evaluated for their sensitivity in detecting Wolbachia in three spider mites, Panonychus mori Yokoyama, Oligonychus gotohi Ehara and Tetranychus kanzawai Kishida. Pure 100% infected populations of these three spider mites were prepared, and female adults from the infected lines at day five were examined for Wolbachia detection using the following PCR primer pairs: wsp, 16S, ftsZ(Bf-Br), ftsZ(f1-r1), ftsZ(Adf-Adr) and ftsZ(Z1-Z2). The wsp primer pair was the most sensitive: it detected the Wolbachia even from a 1/1,000 DNA solution of P. mori with a 47.0% success rate. The 16S primer pair was the second most sensitive. The primer pair ftsZ(Bf-Br) was less sensitive. They were followed by ftsZ(Z1-Z2) and ftsZ(f1-r1) in sensitivity. Thus, the wsp and 16S primer pairs are most suitable for detecting Wolbachia in spider mites, and it is desirable to use these two primer pairs simultaneously for amplifying Wolbachia DNA from single individuals.