Seroreactivity and immunogenicity of Tp0965, a hypothetical membrane protein of Treponema pallidum / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Treponema pallidum (T. pallidum) subsp. pallidum is the causative agent of syphilis. Analysis of recombinant antigens of T. pallidum led to the identification of potential candidate antigens for vaccine development and syphilis serodiagnosis. Tp0965 was predicted to be a membrane fusion protein and was found to be reactive with infected human sera in previous studies, but the results were controversial. In this research, the antigenicity and immunoreactivity of recombinant protein Tp0965 were assessed.</p><p><b>METHODS</b>T. pallidum subsp. pallidum (Nichols strain) was propagated and isolated and the genomic DNA was extracted. The Tp0965 gene was amplified by polymerase chain reaction (PCR). Then the recombinant protein Tp0965 was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid (Ni-NTA) purification system. The reactivities of protein Tp0965 were examined by immunoblot analysis and indirect enzyme-linked immunosorbent assay. The antisera against protein Tp0965 were obtained by immune rabbits and the immunogenicity of antisera were detected by indirect enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>Recombinant protein Tp0965 was expressed successfully in vitro. Immunoblot assay showed that the recombinant protein Tp0965 could be recognized by human syphilitic sera of all stages. Indirect enzyme-linked immunosorbent assay showed there were only 4 of 74 human syphilitic sera that failed to show reactivity to recombinant antigen Tp0965, and lack of reactivity of Tp0965 to all 28 uninfected sera. A low titer of antiserum against Tp0965 in immune rabbits could be detected after the third time of immunization.</p><p><b>CONCLUSIONS</b>The recombinant antigen Tp0965 shows excellent sensitivity for the reactivity with sera from syphilitic individuals at all stages. The results also demonstrate a potential application for the serodiagnosis of syphilis.</p>

Comparative proteome analysis of human papillomavirus-infected cervical specimens and the difference between the high- and low-risk genotypes of human papillomavirus / 中国医学科学院学报

<p><b>OBJECTIVE</b>To perform an comparative proteome analysis of human papillomavirus-infected cervical specimens and to investigate different expressions between high- and low-risk genotypes.</p><p><b>METHODS</b>The cervical specimens were divided into two groups (cervical intraepithelial neoplasia group and condyloma acuminatum group) according to their genotypes. Using comparative proteome technology, high-risk human papillomavirus-infected cervical intraepithelial neoplasia, low-risk human papillomavirus-infected condyloma acuminatum, and normal cervical intraepithelial tissue were compared. The differential expression protein spots were identified by mass spectrometry.</p><p><b>RESULTS</b>Totally 26 differential spots were selected and analyzed, and 22 peptide mass fingerprints (PMF) maps were obtained by MALDI-TOF-MS. Eighteen proteins were preliminarily identified after searching the NCBInr database. The function information of these 18 proteins mainly involved cell metabolism, signal transduction, cell secretion, cell cytoskeleton construction, cell proliferation, and apoptosis.</p><p><b>CONCLUSION</b>The proteomic expressions after the cervical infection of high- or low-risk genotype of human papillomavirus are obviously different.</p>

Genotyping of genital human papillomavirus by DNA sequencing and luminex methods / 中国医学科学院学报

<p><b>OBJECTIVE</b>To compare the specificity and sensitivity of two genotyping approaches for human papillomavirus (HPV).</p><p><b>METHOD</b>HPV DNA was amplified and detected in clinical specimens by polymerase chain reaction in a pair of universal primers MY09/11, and then genotyped with either sequencing method or liquid chip hybridization method (luminex method).</p><p><b>RESULT</b>Sequencing method obtained precise genotyping results in single-type HPV infection, while luminex method obtained accurate genotyping results in multiple-type HPV infection.</p><p><b>CONCLUSION</b>A combined method using both sequencing and luminex method is suitable for the genotyping of HPV-infected specimens.</p>

Epidemiological and bacteriological characteristics of Neisseria gonorrhoeae isolates in China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the antimicrobial susceptibility,auxotype, and plasmid profile of Neisseria gonorrhoeae isolates in China and to provide evidence for the development of treatment guideline and policy for control.</p><p><b>METHODS</b>Agar dilution was used to detect antimicrobial susceptibility. The auxotype was determined by GC genetic medium. The plasmid was extracted by alkaline cleavage and electrophoresed.</p><p><b>RESULTS</b>A total of 4,976 gonococcal isolates were tested in the last 8 years. The resistant rate for penicillin was 71.60% with PPNG being 15.54%. Tetracycline-resistant (TRNG) isolates accounted for 93.02% with 10.48% high level tetracycline-resistant. The resistant rate for ciprofloxacin was also relatively high (31.78%). The resistant rates for spectinomycin and ceftriaxone were 0.36% and 0.46%. The predominant auxotypes of gonococcal isolates were proto and pro(-) during 1995 - 1996 in Nanjing, accounted for 46.4% and 47.53%, 48.4% and 50.22%, respectively. There were 8 strains harboring 4.2, 5.4, 39.5 kb plasmids and 2 harboring 4.2, 4.9, 5.4, 39.5 kb plasmids in 10 PPNG strains; 2 harboring no plasmid, 28 harboring 4.2, 4.9, 5.4, 39.5 kb plasmids in 30 non-PPNG strains. The 5.4 kb plasmid of PPNG could be digested with restriction endonuclease BamHI while the 5.4 kb plasmid of non-PPNG could not.</p><p><b>CONCLUSION</b>The gonococcal isolates were highly resistant to penicillin, tetracycline, and ciprofloxacin, while were still sensitive to spectinomycin and ceftriaxone. No significant auxotyping change was found in terms of predominant gonococcal strains in the last two years in Nanjing while 5.4 kb plasmid might be the most prevalent resistant plasmid in Nanjing.</p>