Detection of human papillomavirus types by the polymerase chain reaction and the differentiation between high-risk and low-risk cervical lesions.

By means of a consensus polymerase chain reaction (PCR) method, the prevalence of HPV types was determined in cervical biopsies from 137 women referred to the gynecological outpatient clinic for colposcopy because of an abnormal cervical smear. The prevalence of HPV was 80.3%. There was a statistically highly significant rise in the prevalence of the oncogenic HPV types (16, 18, 31, 33) with increasing severity of cervical intraepithelial neoplasia (CIN I to III), indicating a role for these HPV types in the pathogenesis of cervical cancer. The prevalence of other HPV types decreased significantly with the severity of the lesion, suggesting that these HPV types play a less significant role in this process. These data indicate that HPV typing with PCR may be a valuable tool for distinguishing between high-risk and low-risk cervical lesions. Furthermore, our results suggest that the detection of HPV types by consensus PCR in the cervix of patients with an abnormal smear but without histologically detectable CIN is a useful tool for predicting which of these patients will eventually develop CIN. Finally, a relatively low percentage (3%) of HPV double infections is reported in this study.     

Significance of high-risk human papillomavirus detection by polymerase chain reaction in primary cervical cancer screening

The purposes of this study were to evaluate the incidence of high-risk human papillomavirus (HPV) infection by polymerase chain reaction (PCR) and to assess its diagnostic usefulness in primary cervical screening. PCR testing for HPV type 16, 18, 31 and 33 was performed on 1305 specimens obtained during routine cervical cancer screening. We analysed the concurrent cervical smears and biopsy, and correlated them with the HPV infection status. We also evaluated histologically-proven cases with ASCUS smears according to HPV infection. HPV DNA was identified in eight (0.7%) of 1144 cytologically normal patients; nine (10.5%) of 86 ASCUS; seven (25.0%) of 28 LSIL; 26 (78.8%) of 33 HSIL; and in all of three squamous cell carcinomas (SCC). HPV positivity was significantly associated with cytohistological diagnosis for HSIL of more. In addition, HPV-positive ASCUS cases were found to be associated with histological abnormality rather than HPV-negative. The results indicate that high-risk HPV testing by PCR could be a useful adjunct tool for Pap smear in primary cervical screening. The combination of Pap smear and high-risk HPV testing by PCR might reduce unnecessary colposcopy-guided biopsy of women with cytological diagnosis of ASCUS.     

阴道微生态改变与人乳头瘤病毒感染和宫颈病变相关研究

目的 探讨阴道微生态变化对人乳头瘤病毒(Human papillomavirus, HPV)感染和宫颈病变的影响。 方法 收集我院2018年11月至2019年7月门诊患者1 529例,进行阴道微生态和宫颈HPV检测,根据筛查结果阴道镜下多点活检,应用卡方检验分析阴道微生态变化在HPV阴性组和阳性组之间差异以及对宫颈病变程度的影响。 结果 pH值>4.6、白细胞计数增高、清洁度级别增高、乳杆菌消失、BV和滴虫感染能增加HPV感染(χ2值分别为12.281、17.692、79.999、192.700、48.863和16.758,均P结论 阴道微生态变化能增加HPV感染概率,但与HPV相关性宫颈病变关系不密切。     

人细小病毒B19与自然流产关系的研究

无菌留取 5 4例自然流产妇女和 43例妊娠无异常孕妇血清 ,用聚合酶链反应 (PolymeraseChainReaction ,PCR)检测的人细小病毒B19(HumanParvovirusB19,B19)DNA ,在自然流产组中人细小病毒B19DNA有 15例阳性 ,阳性率为 2 7.78%。正常对照组中 ,人细小病毒B19DNA有 2例为阳性 ,阳性率为 4.65 % ,用x2 检验 ,x2 =8.86,P <0 .0 1,两组有非常显著性差异。由此总结 ,人细小病毒B19感染可能是导致自然流产的原因之一     

Detection by multiplex polymerase chain reaction and typing of Chlamydia trachomatis isolates

Abstract The multiplex polymerase chain reaction (PCR) was applied for the detection of the Chlamydia trachomatis chromosome and plasmid. The multiplex PCR demonstrated a sensitivity of 0.8 fg of chlamydial DNA, corresponding to the detection of about 5 copies of the plasmid. Analysis of 195 genital specimens collected randomly from a female population, showed that the multiplex PCR is more sensitive and rapid than culturing for detecting Chlamydia trachomatis . Moreover, sequencing of the II variable domain of the ompl gene, directly from DNA of the clinical specimens, appears to be a simple and rapid method for determining serovar isolates.     

STAT3基因多态性与女性人乳头瘤病毒感染及宫颈病变的关系

目的探究女性人乳头瘤病毒(HPV)感染及转录活化因子3(STAT3)单核苷酸基因多态性(SNPs)的改变。方法2016年9月至2019年9月经病理组织学确定的健康子宫者60例(NC组)、低度鳞状上皮内病变(LSIL)50例(LSIL组)、高度鳞状上皮内病变(HSIL)55例(HSIL组)和宫颈鳞状细胞癌(CSCC)57例(CSCC组),PCR测定HPV感染情况,同时采用PCR 限制片段长度多态性(RFLP)测定各组STAT3基因C1697G多态性。结果HSIL组、LSIL组及CSCC组高危HPV与NC组比较阳性率显著升高,CSCC组(98.24%)显著高于HSIL组(72.73%)和LSIL组(5.8%),HSIL组显著高于LSIL组,结果均具有统计学差异(P<0.05)。LSIL组、HSIL组及CSCC组中与NC组比较显著升高的基因比例为C/C型,CSCC组C/C型基因比例为70.18%,HSIL组(58.18%)C/C型基因比例显著高于LSIL组(0.18%)(P<0.05)。与HPV阴性比较,C/C型基因比例在HPV阳性中显著升高(P<0.05)。通过调整年龄因素,C/G基因型HPV感染风险上升到3.120(95％CI:0.847～4.995),发生宫颈癌的风险上升到3.876(95％CI为0.935～5.336);C/C型基因HPV感染风险上升到3.574(95％CI为0.893～4.912),发生宫颈癌的风险上升到5.137(95％CI为0.967～5.776)。结论STAT3基因C1697G多态性与HPV易感性及宫颈病变的发生发展有密切关系。     

人乳头瘤病毒感染与宫颈癌的研究进展

人乳头瘤病毒(Human Papillomavirus HPV)感染是导致性传播疾病的常见原因,上世纪八十年代初,首次报道生殖器HPV感染与宫颈癌之间的联系,认为HPV感染是95%以上宫颈癌变的高危因素。随着分子生物学技术的发展,对HPV致癌机制的研究不断深入,取得大量有价值的成果,现就HPV的致癌途径与协同因素探讨宫颈癌的发病机制以及对HPV检测方法等方面的研究进行综述。     

Detection of Norwalk virus in the UK by the polymerase chain reaction

Abstract We have developed a polymerase chain reaction for the detection of Norwalk virus using the published sequence of the virus RNA dependent RNA polymerase gene and have used this to clone and sequence this region of a virus from a UK outbreak. We have applied this method to a panel of UK Norwalk-like viruses using both Tet-z and Taq DNA polymerases and found that amplification produces a multiplicity of bands from stool samples. However, combination with Southern blotting. Taq polymerase amplification detected virus in 13 of a panel of 30 clinical samples known to contain these viruses and also detected astroviruses in a mixed infection. Amplification using Tet-z DNA polymerase was less efficient (6/30) and detected predominantly viruses typed as UK type 2 by solid phase immune electron microscopy.     

A retrospective analysis of human papillomavirus (HPV) prevalence and genotype distribution among 25,238 women in Shanghai,China revealed the limitations of current HPV-based screening and HPV vaccine

BackgroundTo determine the human papillomavirus (HPV) type-specific prevalence and distribution among women with various age and cervical lesions in Shanghai, China. And to evaluate the carcinogenicity of different high-risk HPV (HR-HPV) and the efficacy of HR-HPV testing and HPV vaccine.MethodsThe clinical data from 25,238 participants who received HR-HPV testing (HPV GenoArray test kit, HybriBio Ltd) at the Affiliated Hospital of Tongji University from 2016 to 2019 were reviewed and analyzed using SPSS (version 20.0, Tongji University, China).ResultsThe overall prevalence of HPV was 45.57% in the study population, of which 93.51% were found HR-HPV infection. The three most prevalent HR-HPV genotypes were HPV 52 (22.47%), 16 (16.4%) and 58 (15.93%) among HPV-positive women, and HPV 16 (43.30%), 18 (9.28%) and 58 (7.22%) in women with histologically confirmed cervical cancer (CC). 8.25% of CC were found to be HPV negative. Only 83.51% of CC cases were related to the HPV genotypes covered by nine-valent HPV vaccine. HPV prevalence and genotype distribution varied with age and cervical histology. The odds ratios (OR) of HR-HPV for CC were also different, among which the top three types were HPV 45 [OR= 40.13, 95% confidence intervals (CI) 10.37–155.38], 16 (OR=33.98, 95%CI 15.90–72.60) and 18 (OR=21.11, 95%CI 8.09–55.09). The increase in the types of HPV infection did not increase the risk of CC correspondingly. As the primary cervical screening method, HR-HPV testing showed the high sensitivity (93.97%, 95%CI 92.00–95.49) but low specificity (42.82%, 95%CI 41.81–43.84).ConclusionsOur study provide the comprehensive epidemiological data on HPV prevalence and genotype distribution among Shanghai women with various cervical histology, which can not only serve as a significant reference for clinical practice, but also implicated the need of more effective CC screening methods and HPV vaccine covering more subtypes.     

阴道微生态异常与高危型人乳头瘤病毒感染及宫颈病变的关系

目的 探讨阴道微生态异常与高危型人乳头瘤病毒（HR-HPV）感染及宫颈病变的相关性。方法 选取2015年1月至2017年6月在十堰市妇幼保健院妇科门诊接受宫颈病变筛查的562例患者为研究对象,分别进行阴道微生态、HR-HPV检测、宫颈脱落细胞检查和组织病理学活检,对检查结果进行分析。结果 562例患者中,宫颈病变发生率为16.01%,液基薄层细胞检测（TCT）阳性率为19.93%,HR-HPV感染率为28.47%,阴道微生态失调发生率为30.78%,其中细菌性阴道病（BV）、滴虫性阴道炎（TV）、外阴阴道假丝酵母菌病（VVC）、需氧菌性阴道炎（AV）和乳杆菌减少检出率分别为24.02%、3.20%、12.28%、8.54%和28.47%。宫颈病变患者微生态失调率为67.78%,明显高于正常组的23.73%（P<0.05）。宫颈病变患者其BV、VVC、AV和乳杆菌减少检出率分别为66.67%、30.0%、22.22%和75.56%,均明显高于正常者的15.89%、8.90%、5.93%和19.49%（P<0.05）;宫颈病变患者HR-HPV阳性率为75.56%,明显高于正常者的19.49%（P<0.05）。HR-HPV阳性患者BV、VVC、AV和乳杆菌减少的检出率分别为51.88%、24.38%、16.88%和62.5%,明显高于HR-HPV阴性患者的12.94%、7.46%、5.22%和14.93%（P<0.05）。结论 阴道微生态环境的改变与高危型HPV感染及宫颈病变的发生具有一定关系。     

Detection of Chlamydia psittaci using DNA probes and the polymerase chain reaction

Fewer than 10(5) elementary bodies of Chlamydia psittaci could be detected by using DNA hybridisation with a plasmid probe specific for avian chlamydial strains. PCR amplification of chlamydial DNA using primers specific for conserved regions of the major outer membrane protein gene enabled the detection of fewer than 10 elementary bodies. DNA could be amplified from 22 of the 24 chlamydial strains tested including avian, feline, ovine, caprine, koala and lymphogranuloma venereum strains.     

Detection of Nitrosomonas spp. by polymerase chain reaction

Abstract A unique genomic DNA fragment was isolated from Nitrosomonas europaea ATCC 19718. Based on the sequence of this fragment, oligonucleotide primers for polymerase chain reaction amplification were prepared which amplify sequences of 775 and 658 bp. The predicted DNA fragments were both amplified from the genome of N. europaea and a Nitrosomonas spp. isolated from a local oxidation pond. The primers failed to amplify DNA from the genomes of the ammonia oxidiser Nitrosolobous multiformis , the nitrite oxidiser Nitrococcus mobilis as well as from the genomes of other unrelated heterotrophic bacteria. These DNA sequences could be amplified from 0.01 ng of N. europaea genomic DNA or from 100 intact cells, and it was possible to detect Nitrosomonas DNA in a DNA mixture extracted from water samples drawn from a local oxidation pond.     

宫颈上皮内瘤变患者阴道微生态状况与LEEP术后预后关系

目的 研究宫颈上皮内瘤变（CIN）患者阴道微生态状况及LEEP术后阴道微生态状况的变化特点,分析阴道微生态状况与CIN患者预后的关系。方法 回顾性分析2016年1月至2018年1月温州医科大学定理临床学院122例高危型人乳头瘤病毒（hr-HPV）阳性的CIN患者LEEP术治疗前后阴道微生态的变化特点。结果 LEEP术后菌群密集度Ⅱ~Ⅲ级（74.59%）、多样性Ⅱ~Ⅲ级（68.03%）、乳酸分级Ⅰ~Ⅱa级（70.49%）出现增多,需氧菌性阴道病（AV）（9.84%）、细菌性阴道病（BV）（14.75%）、外阴阴道假丝酵母菌病（VVC）（12.30%）以及滴虫性阴道炎（2.46%）的检出率降低和pH值（4.2±0.5）降低,与术前比较差异具有统计学意义（均P<0.05）;术后复诊hr-HPV阳性CIN患者菌群密集度Ⅱ~Ⅲ级（35.00%）、多样性Ⅱ~Ⅲ级（40.00%）、乳酸分级Ⅰ~Ⅱa级（25.00%）低于hr-HPV阴性CIN患者,AV（30.00%）、BV（35.00%）、VVC（30.00%）检出率,pH值（4.9±0.4）高于hr-HPV阴性CIN患者,差异有统计学意义（均P<0.05）。结论 阴道微生态失衡可降低宫颈免疫力,促进CIN进展、癌变,改善阴道微生态可抑制hr-HPV的感染,抑制宫颈细胞的异常增生、癌变。     

CIN患者阴道微生态状况与HPV感染的关系分析

分析宫颈上皮内瘤变（CIN）患者阴道微生态与人乳头瘤病毒（HPV）感染的关系。

选取326例本院收治的CIN患者,采集阴道分泌物和宫颈脱落细胞,通过形态学和功能学检测阴道微生态状况,采用PCR-反向点杂交法进行HPV分型检测,采用二分类Logistic回归分析影响CIN患者HPV感染的因素。

CIN患者HPV阳性共256例,占78.5%,HPV阴性共70例,占21.5%。HPV阳性CIN患者白细胞计数＞10个/高倍视野、阴道清洁度Ⅲ-Ⅳ度、VVC阳性、AV阳性、BV阳性、pH值＞4.5、白细胞酯酶阳性、过氧化氢阳性占比高于HPV阴性患者,微生态正常者占比低于HPV阴性患者（P＜0.05）。二分类Logistic回归分析显示,VVC、BV、微生态正常和pH值为CIN患者HPV感染的影响因素。

CIN患者HPV感染率较高,且感染后存在阴道微生态失调,VVC、BV、微生态异常和pH值是CIN患者HPV感染的影响因素。

     

Human papillomavirus genotype distribution in cervical intraepithelial neoplasia grades 1 or worse among 4215 Chinese women in a population-based study

PurposeTo estimate the burden of human papillomavirus (HPV) infection and cervical disease among sexually active women in a sample of Chinese women.MethodsA multicenter, population-based study was conducted between May 2006 and April 2007. A total of 4215 sexually active women aged 17–54 years were surveyed from five geographical sites representing both urban and rural areas: Beijing, Shanghai, Shanxi, Henan and Xinjiang. Women were referred for colposcopy on the basis of results of Pap testing and HPV screening. HPV genotyping of the CIN1+ specimens was performed with INNO-LiPA. Attribution of HPV types to lesions was estimated using a fractional contribution approach.Results13.3% of the women (559/4215) were referred for colposcopy; 4.3% (183/4215) of these were diagnosed with CIN1+. Of the latter, 88.5% (162/183) were typed and 94.4% (153/162) were HPV-positive. HPV16 was the most prevalent type in lesions in both urban and rural settings. Combined, HPV16 and 18 were attributable to 71.4% of HPV-positive CIN2+ lesions. In addition, HPV31, 33, 52 and 58 were prevalent in CIN1+ lesions, with HPV33, 52, and 58 combined accounting for 24.1% CIN2+ lesions. Though prevalent, HPV31 always occurred as a co-infection with another HPV type and therefore was attributed minimal causality.ConclusionsHPV16 and 18 are associated with the majority of cervical lesions in Chinese women from which this population-based sample was drawn. In addition, other HPV types, such as 33, 52, and 58, also play an important role in cervical disease.     

Detection of urovirulence factors in Escherichia coli by multiplex polymerase chain reaction

Abstract Primers to amplify the genes encoding the virulence factors of uropathogenic Escherichia coli , such as pilus associated with pyelonephritis ( pap ), haemolysin ( hly ), aerobactin ( aer ) and cytotoxic necrotizing factor 1 ( cnf 1) genes, were designed. The above primers along with previously reported primers for S fimbriae ( sfa ) and afimbrial adhesin I ( afaI ) genes were combined to develop a multiplex polymerase chain reaction (PCR) for detection of the respective virulence factors and for the identification of uropathogenic E. coli . The multiplex PCR to detect pap, sfa, afa I, hly, aer and cnf 1 genes was highly specific and the sensitivity was found to be about 5 × 10³ colony forming units of E. coli per ml. A total of 194 E. coli strains isolated from patients with simple acute cystitis were examined by the multiplex PCR and the results were in complete agreement with that obtained by DNA colony hybridization test. The multiplex PCR developed was, therefore, concluded to be a useful, sensitive and rapid assay system to identify uropathogenic E. coli .     

Detection of Mycobacterium leprae in nasal mucosa biopsies by the polymerase chain reaction

Several discoveries about leprosy indicate that Mycobacterium leprae transmission mainly occurs by inhalation, and the nose is major port of entry and exit. The present study evaluated the clinical application of PCR for detection of M. leprae DNA in nasal mucosa biopsies in untreated leprosy patients (52) and their contacts (99) from the State Reference Center in Sanitary Dermatology and Leprosy, Uberlandia, MG, Brazil. PCR detection of a 372-base pair DNA fragment from M. leprae was accomplished in 36 (69.2%) patients, from which 34 (91.9%) of them were multibacillaries. Furthermore, PCR was positive in 3 (16.7%) of 18 slit-skin smear negative, 4 (25.0%) of 16 skin lesion BI negative, 8 (33.3%) of 24 nasal mucosa BI negative patients, and 10 of 99 contacts (10.1%). The presence of bacilli in 10.1% of the contacts may potentially reflect an occult leprosy, and these patients must be accompanied, followed by a chemoprophylaxy treatment. Considering all PCR results against clinical and BI classification of patients and controls, we have found a sensitivity of 69.2%, a specificity of 89.9%, and an accuracy of 82.8%. It has been demonstrated here through PCR of nasal biopsies that the bacillus invades the mucosa, passing through the nasal inferior turbinate to reach peripheral blood. Therefore, the molecular investigation of invasive nasal biopsies by PCR tests has proven to be useful in defining patients of higher risk of transmission and risk-group contacts, which is an important step to reach the World Health Organization objective towards the elimination of leprosy as a public health problem.     

Neutralizing antibodies against oncogenic human papillomavirus as a possible determinant of the fate of low-grade cervical intraepithelial neoplasia

To determine whether neutralizing antibodies (NAs) against HPV16 is responsible for a higher regression rate of low-grade cervical intraepithelial neoplasia (CIN1), we investigated an association between the presence of the NAs and the fate of the HPV16-related CIN1. All the women examined in this study had HPV16 positive cervix. The women were allocated into four groups by their cervical pathology, i.e., non-pathological (n:7), CIN1 (n:37), CIN2/3 (n:19), and cervical cancer (n:13). Their sera were tested for the presence of NAs against HPV16 by an in vitro assay using HPV16-pseudovirions. As for the CIN1 cases, clinical regression of the lesions were compared between NA-positive and NA-negative groups. Copy number of HPV16-DNA in smear samples was measured by quantitative PCR. The incidence of the presence of the NAs in the women with a non-pathological cervix (85.7%) was significantly higher than in the CIN1 cases (21.5%), the CIN2/3 cases (15.7%), and the cervical cancer cases (0%) (p<0.0001). The regression of the CIN1 lesion was closely associated with the presence of the N As (p=0.0002). The presence of the NAs was associated with low-level copy number of the viral DNA relative to the NA-negative group (p=0.05). The presence of the NAs against HPV16 was associated with a higher regression rate of HPV-related CIN1 lesions. The NAs seem to have a role in deterring HPV-related cervical lesions from progressing to CIN2/3 by inhibiting the infection with de novo replicated HPV. This study further suggests that HPV vaccine to induce the NAs may be effective in eliminating CIN lesions, especially in the NA-negative cases.     

伴高危型HPV感染的LSIL患者HPV清除与 阴道微生态间的关系

目的探讨阴道微生态对伴有高危型人乳头瘤病毒(HR-HPV)感染的低级别鳞状上皮内病变(LSIL)患者HPV清除速度有无影响。方法选取阴道镜下宫颈活检病理诊断为LSIL或CIN1,且均合并HR-HPV感染的患者351例,随访1～2年,观察入组时的阴道微生态状态与随访6个月及12个月时的HPV清除情况。结果对于有随访结果的334例,至随访6个月时,细菌性阴道病(BV)组HPV转阴率为20.29%,低于阴道微生态正常组(32.35%,P0.01),其余外阴阴道假丝酵母菌组、滴虫性阴道炎组及其他微生态失衡组与阴道微生态正常组间比较差异无统计学意义。至随访12个月时,各微生态失衡组与微生态正常组间HPV转阴率差异均无统计学意义。结论 BV感染可能在短期内影响LSIL患者HPV清除速度。