Improvement on conditions of PCR/SSCP and its application in rapid genotyping of orientia tsutsugamushi

  Objective   In order to rapidly analyze the gene type of orientia tsutsugamushi(Ot)from Shandong province by PCR/SSCP.

  Methods   Comparing the SSCP maps of the amplification products by group primers of Ot-DNA acquired from 23 strains of Ot isolated, 2 pools of homogenized Leptotrombidium(L.)scutellare, and 10 blood of scrub typhus with the reference strains Gilliam、Karp、Kato, and also compared with the results acquired by Nested PCR.

  Results   Ot-Sta56 gene amplification fragments by group primers of Ot-DNA acquired from 23 strains of Ot isolated, 2 pools of homogenized L.scutellare, and 10 blood of scrub typhus appeared two different maps of SSCP, 33 had the same gene types different from that of the reference strain Gilliam、Karp、Kato, 2 had the same gene type with the reference strain Karp.This results were identical to that acquired by Nested PCR.

  Conclusion   PCR/SSCP was a rapid, simple, accurate method for analyzing the gene type of Ot.