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目的:通过对正常孕妇及胎儿HPV感染状况的研究,分析HPV宫内感染可能存在的途径。方法:以GP5+/6+为引物的通用引物PCR(GP-PCR),被证明是一种敏感性较高的人乳头瘤病毒检测方法〖16、18、19〗。本研究在此方法的基础上,利用生物素标记的引物(bio-GPbt),进行GP-PCR,并用因此而产生的生物素标记的PCR产物与特异性内寡核苷酸探针(SIOP)进行反向点杂交(RDBH),从而达到检测HPV感染并对其分型的目的。结果:1.在总计119份样品中,母血、胎儿脐带血、羊水、宫颈分泌物、胎盘组织母体面的HPV感染率分别为37.0%(10/27)、37.0%(10/27)、61.11%(11/18)、50.0%(10/20)、40.7%(11/27),说明妊娠末期孕妇有较高的感染率;2.对取自同一标本的5种样品(母血、胎儿脐带血、羊水、宫颈分泌物、胎盘组织母体面)进行检测,由于5种样品之间的感染率无显著性差别(P>0.1),结合观察到的实际结果,推测HPV有通过宫内感染途径感染胎儿的可能性;3.本研究还发现,在52份HPV阳性的样品中,有12份(23.1%)为多重HPV感染。另外,52份HPV阳性的样品(100%)均发现HPV11型,推测妊娠末期的孕妇主要以HPV11型感染为主。
OBJECTIVE: To analyze the possible ways of HPV intrauterine infection by studying the status of HPV infection in normal pregnant women and fetuses. Methods: The universal primer PCR (GP-PCR) using GP5 + / 6 + as a primer has been proved to be a highly sensitive method for the detection of human papillomavirus [16,18,19]. In this study, based on this method, GP-PCR was performed using biotin-labeled primers (bio-GPbt), and the resulting biotin-labeled PCR product and specific internal oligonucleotide probe (SIOP) Reverse dot blot (RDBH) was performed to detect and classify HPV infection. HPV infection rates of maternal blood, fetal umbilical cord blood, amniotic fluid, cervical secretions and placenta tissue surface were found to be 37.0% (10/27) and 37.0% (10/27) respectively in a total of 119 samples. 61.11% (11/18), 50.0% (10/20), 40.7% (11/27), indicating that pregnant women have a higher infection rate at the end of pregnancy. (2) Five samples collected from the same sample Fetus umbilical cord blood, amniotic fluid, cervical secretions, placenta tissue mother face), the infection rate of the five samples was no significant difference (P> 0.1), combined with the observed actual results, it is speculated that HPV through intrauterine The possibility of infecting the fetus by the route of infection; 3. The study also found that of the 52 HPV-positive samples, 12 (23.1%) were multiple HPV infections. In addition, 52 HPV-positive samples (100%) were found HPV11 type, presumably pregnant women in the end of pregnancy mainly HPV11-based infection.