CAJ | 학술논문

目的：建立微小脲原体（ Up）聚合酶链反应（ PCR）相对定量方法，克服阴道标本采样差异对检测结果的影响，并探讨Up在细菌性阴道病（ BV）中的作用。方法根据Up 4个血清型拓扑异构酶Ⅳ基因序列，合成Up定量PCR的引物和探针，建立Up PCR定量方法；同时进行宿主细胞甘油醛-3-磷酸脱氢酶（ GAPDH）基因定量，计算103宿主细胞中Up的相对含量；分别检测Up标准株和临床分离株、解脲脲原体标准株和临床分离株、11种其它阴道常见微生物，验证PCR的特异性；从532例BV患者和276名健康女性采集阴道分泌物，进行Up相对定量分析。结果 Up定量PCR方法能直接检测阴道分泌物中的Up，对解脲脲原体和11种其它阴道常见微生物无交叉反应，灵敏度为100拷贝／μL。 BV患者和健康女性的Up感染率分别为49．8％（265／532）和43．1％（119／276），二者之间差异无统计学意义（χ2＝3．263，P＝0．071）；BV患者和健康女性Up相对定量分别为667拷贝／103细胞和20拷贝／103细胞，二者比较差异有统计学意义（χ2＝47．012，P＝0．0001）。如果以50拷贝／103细胞作为参考值，则BV患者Up的阳性率为39．3％（209／532），健康女性的阳性率为17．0％（47／276），二者比较差异有统计学意义（χ2＝41．537，P＝0．001）。结论 Up黏附到宿主细胞表面是其致病的关键因素。建立的Up相对定量方法能反映宿主细胞上Up的含量，克服了采样差异对检测结果的影响，能更客观地反映Up的致病能力。
목적：건립미소뇨원체（ Up）취합매련반응（ PCR）상대정량방법，극복음도표본채양차이대검측결과적영향，병탐토Up재세균성음도병（ BV）중적작용。방법근거Up 4개혈청형탁복이구매Ⅳ기인서렬，합성Up정량PCR적인물화탐침，건립Up PCR정량방법；동시진행숙주세포감유철-3-린산탈경매（ GAPDH）기인정량，계산103숙주세포중Up적상대함량；분별검측Up표준주화림상분리주、해뇨뇨원체표준주화림상분리주、11충기타음도상견미생물，험증PCR적특이성；종532례BV환자화276명건강녀성채집음도분비물，진행Up상대정량분석。결과 Up정량PCR방법능직접검측음도분비물중적Up，대해뇨뇨원체화11충기타음도상견미생물무교차반응，령민도위100고패／μL。 BV환자화건강녀성적Up감염솔분별위49．8％（265／532）화43．1％（119／276），이자지간차이무통계학의의（χ2＝3．263，P＝0．071）；BV환자화건강녀성Up상대정량분별위667고패／103세포화20고패／103세포，이자비교차이유통계학의의（χ2＝47．012，P＝0．0001）。여과이50고패／103세포작위삼고치，칙BV환자Up적양성솔위39．3％（209／532），건강녀성적양성솔위17．0％（47／276），이자비교차이유통계학의의（χ2＝41．537，P＝0．001）。결론 Up점부도숙주세포표면시기치병적관건인소。건립적Up상대정량방법능반영숙주세포상Up적함량，극복료채양차이대검측결과적영향，능경객관지반영Up적치병능력。
Objective To establish Ureaplasma parvum ( Up ) polymerase chain reaction ( PCR ) relative quantitative assay, to avoid the effect of different vaginal sampling ways for determination results, and to investigate the role of Up in bacterial vaginosis ( BV) .Methods The primers and probes for Up were designed according to DNA topoisomerase Ⅳ gene sequences of 4 Up serovars to detect Up in vaginal secretion samples.Glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) gene was analyzed by PCR quantitative assay to determine host cells.The Up relative quantification was calculated as Up copies in 103 host cells.Up standard isolate and clinical isolates, Ureaplasma urealyticum standard isolate and clinical isolates and 11 common vaginal microorganisms were determined to identify the specificity of the assay.The vaginal secretion samples were collected from 532 BV patients and 276 healthy women for the analysis of Up relative quantification. Results Up PCR quantitative assay had high sensitivity (100 copies/μL) , without amplification to Ureaplasma urealyticum and 11 other vaginal microorganisms.The infection rates of Up in BV patients and healthy women were 49.8%(265/532) and 43.1%(119/267), respectively, without statistical significance (χ2 =3.263, P=0.071).Up relative quantification in BV patients was 667 copies/103 cells, which was significantly higher than that in healthy women (20 copies/103 cells) (χ2 =47.012,P=0.000 1).When using >50 copies/103 cells as a reference, the positive rate of Up in BV patients was 39.3% (209/532).It was significantly higher than that in healthy women (17.0%, 47/276) (χ2 =41.537,P=0.001).Conclusions The cell adhesion ability of Up is a key factor in the pathogenesis.Up PCR relative quantitative assay can overcome the influence of different vaginal sampling ways on the determination results and objectively reflect the pathogenicity of Up.