Development Of PCR-CE Method For Detection Of Biovars Of Ureaplasma Urealyticum And Its Application On Etiological Study Of NGU

Background: Ureaplasma urealyticum is a member of genus Ureaplasma,family Mycoplasmataceae, order Mycoplasmatales, class Mollicutes. U.urealyticum are divided into two biovars, biovar 1 and biovar 2. U. urealyticum can cause a number of diseases of human, but the pathogenicity of U. urealyticum in the male with nongonococcal urethritis (NGU) remains controversial. In 1999, biovar 1 were designated as a separate species, Ureaplasma parvum, whereas biovar 2 retained the U.urealyticum designation. Thus, an association between U. urealyticum and NGU should probably be understood in terms of biovars.Up to now, culture has been widely used in detection of U. urealyticum in clinical laboratory, but culture needs longer time and can not distinguish these two biovars. There are some differences in Multiple Banded antigen gene(mba gene),16S rRNA gene, 16S-23S rRNA gene and urease gene of two biovars of U. urealyticum, which can be detected by polymerase chain reaction(PCR). Capillary electrophoresis(CE) is a rapid, sensitive, high-powered automated technique with higher resolution and has been used in detection of nucleotides. At present, PCR-CE has been widely used in microbiological detection.Objective:The objective of this study is to develop a PCR-CE method for detection of two biovars of U. urealyticum and to detect two biovars of U. urealyticum in urine from different population using this method in order to determine whether two biovars of U. urealyticum are associated with NGU of the male.Methods: Primers UMS-125,UMA-226 located at mba gene of U. urealyticum were synthesized. Primer UMS-125 was 5'-labeled with FAM. Two biovars of U. urealyticum were amplified by PCR with the two primers . Based on the trivial length difference of the PCR products discriminated by CE, the two biovars can be...