A multicenter prospective trial to asses a new real-time polymerase chain reaction for detection of Treponema pallidum,herpes simplex-1/2 and Haemophilus ducreyi in genital,anal and oropharyngeal ulcers

Treponema pallidum, herpes simplex virus types 1 or 2 (HSV-1/2) and Haemophilus ducreyi are sexually transmitted pathogens that can cause genital, anal and oropharyngeal ulcers. Laboratory evaluation of these pathogens in ulcers requires different types of specimens and tests, increasing the risk of improper specimen handling and time lapse until analysis. We sought to develop a new real-time PCR (TP-HD-HSV1/2 PCR) to facilitate the detection of T. pallidum, HSV-1/2 and H. ducreyi in ulcers. The TP-HD-HSV1/2 PCR was tested (i) in a retrospective study on 193 specimens of various clinical origin and (ii) in a prospective study on 36 patients with genital, anal or oropharyngeal ulcers (ClinicalTrials.gov # NCT01688258). The results of the TP-HD-HSV1/2 PCR were compared with standard diagnostic methods (T. pallidum: serology, dark field microscopy; HSV-1/2: PCR; H. ducreyi: cultivation). Sensitivity and specificity of the TP-HD-HSV1/2 PCR for T. pallidum were both 100%, for HSV-1 100% and 98%, and for HSV-2 100% and 98%, respectively. T. pallidum and HSV-1/2 were detected in 53% and 22% of patients in the prospective study; H. ducreyi was not detected. In the prospective study, 5/19 (26%) specimens were true positive for T. pallidum in the TP-HD-HSV1/2 PCR but non-reactive in the VDRL. The TP-HD-HSV1/2 PCR is sensitive and specific for the detection of T. pallidum and HSV-1/2 in routine clinical practice and it appears superior to serology in early T. pallidum infections.     

Multicentre surveillance of prevalence of the 23S rRNA A2058G and A2059G point mutations and molecular subtypes of Treponema pallidum in Taiwan, 2009–2013

Resistance mutations A2058G and A2059G, within the 23S rRNA gene of Treponema pallidum, have been reported to cause treatment failures in patients receiving azithromycin for syphilis. Genotyping of T. pallidum strains sequentially isolated from patients with recurrent syphilis is rarely performed. From September 2009 to August 2013, we collected 658 clinical specimens from 375 patients who presented with syphilis for genotyping to examine the number of 60-bp repeats in the acidic repeat protein (arp) gene, T. pallidum repeat (tpr) polymorphism, and tp0548 gene, and to detect A2058G and A2059G point mutations by restriction fragment length polymorphism. Treponemal DNA was identified in 45.2% (n = 298) of the specimens that were collected from 216 (57.6%) patients; 268 (40.7%) specimens tested positive for the 23S rRNA gene, and were examined for macrolide resistance. Two isolates (0.7%) harboured the A2058G mutation, and no A2059G mutation was identified. A total of 14 strains of T. pallidum were identified, with 14f/f (57.5%) and 14b/c (10.0%) being the two predominant strains. Forty patients who presented with recurrent episodes of syphilis had T. pallidum DNA identified from the initial and subsequent episodes, with five cases showing strain discrepancies. One patient had two strains identified from different clinical specimens collected in the same episode. Our findings show that 14f/f is the most common T. pallidum strain in Taiwan, where the prevalence of T. pallidum strains that show A2058G or A2059G mutation remains low. Different genotypes of T. pallidum can be identified in patients with recurrent episodes of syphilis.     

A Three Year Retrospective Study on Seroprevalence of Syphilis among Pregnant Women at Gondar University Teaching Hospital,Ethiopia

Background

Sexually transmitted infections (STIs) are a serious public health problem in low income countries, including Ethiopia. Syphilis caused by Treponema pallidum remains a major cause of reproductive morbidity and poor pregnancy outcomes in low income countries. Stillbirth, perinatal death, serious neonatal infection and low-birth weight babies are common among seropositive mothers.

Objective

To assess the seroprevalence of syphilis and risk factor correlates of this infection at Gondar University Teaching Hospital, Ethiopia.

Methods

The study was done on 2385 pregnant women attending the antenatal care clinic (ANC) from January 2009 to December 2011. Data was abstracted from the antenatal care clinic medical database. Chi-square test was used, using SPSS version 16 and significance level was chosen at 0.05 level with a two-tailed test.

Results

Of the total, 69(2. 9%) of pregnant women were confirmed as seropositive for syphilis. Pregnant women with an age group of 21–25 years of old were the most seropositive (3.4%), followed by 26–30 years of old (3.1%). The prevalence of syphilis infection was 3.2% in urban and 2.2% in rural pregnant women. Relatively high prevalence of syphilis infection were identified among students (4.2%) followed by governmental employees (3.8%).

Conclusion

The seroprevalence of syphilis among pregnant women attending ANC is declining. However, syphilis is more prevalent in the young and urban pregnant women. Emphasis on education to young people on STI risk behavioral change and partner follow up and notification for exposure to syphilis and treatment should be given.     

2011年深圳市梅毒母婴垂直传播影响因素分析

目的掌握梅毒在深圳市孕产妇中的感染现状和流行病学特点,为阻断梅毒母婴垂直传播提供参考。方法采用快速血浆反应素环状卡片试验法（RPR）对2011年1-12月深圳市某区的孕产妇进行梅毒检测的初筛。所有阳性血清送往当地疾病预防控制中心进行梅毒螺旋体明胶凝集试验法（TPPA）确诊试验;对阳性孕产妇进行问卷调查和随访分析,并对阳性孕产妇新生儿出生1个月和6个月后进行2次梅毒检测。结果深圳市某区2011年度共有孕产妇16153例,其中普通孕产妇15798例,特殊孕产妇355例,共有15788例接受梅毒血清学检测,检测覆盖率为97．77％,其中普通孕产妇15433（检测覆盖率为97．63％）和特殊孕产妇355例（检测覆盖率为100．00％）。共检测出56例梅毒阳性的孕产妇,其中普通孕产妇中50例,特殊孕产妇中有6例,阳性率为0．35％;共检测出17例先天梅毒新生儿,阳性率为0．11％：多因素Logistic回归分析结果显示：职业和文化程度与孕产妇梅毒感染关系密切。结论深圳市孕产妇梅毒初筛覆盖率较高,孕产妇的职业和文化程度与母婴梅毒垂直传播关系密切。     

Mumbai disease in far western Nepal: HIV infection and syphilis among male migrant-returnees and non-migrants

OBJECTIVE: To measure the seroprevalence of human immunodeficiency virus (HIV) infection and syphilis, and to assess the behavioural risk factors for these infections among migrant-returnees and non-migrants in far western Nepal. METHODS: In April 2001, we recruited 97 male migrant-returnees and 40 non-migrants from five rural villages in Doti district where migration to Mumbai is common. For data collection, we conducted a serological examination for HIV and syphilis, and a perception and behaviour survey on HIV and other sexually transmitted infections. RESULTS: We found that 11 of 137 men (8%) were positive for HIV infection and 30 men (22%) for syphilis. The respondents, especially the migrant-returnees from Mumbai, were engaging in risky behaviours such as pre- or extramarital sex, and sex with multiple partners, including sex workers. CONCLUSIONS: This study revealed high HIV and syphilis prevalence among the male migrant-returnees and non-migrants in far western Nepal where migration to Mumbai is common. The prevalent behaviours, particularly among the migrant-returnees, imply urgent needs of the behavioural modification programme in this area to prevent the spread of HIV infection to general population.     

化学发光法检测受血者梅毒螺旋体特异性抗体的方法学评价

目的 通过与梅毒螺旋体微量血凝试验(TPHA)和蛋白印迹试验法(WB)比较评估梅毒血清学筛查法化学发光法(CLIA)的性能.方法 回顾性研究18 494例受血者血清标本CLIA和TPHA检测梅毒螺旋体特异性抗体结果,对CLIA和(或)TPHA结果阳性177例标本用WB检测确认.同时用CLIA、TPHA和WB检测了81例各期梅毒患者血清、55例有潜在干扰的患者血清和250例阴性对照血清梅毒抗体.结果 以WB检测结果为金标准,CLIA方法的灵敏度为98.4％,显著高于TPHA(94.4％)(x2=5.76,P＜0.05);CLIA方法特异性为100％,高于TPHA方法特异性(99.7％),但差异无统计学意义(x2=1.0,P＞0.05).结论 CLIA法具有高度敏感性和特异性,适合于临床实验室进行梅毒筛查.     

Clinicopathological features of syphilitic gastritis in Korean patients

Syphilitic gastritis is difficult to diagnose without a history of syphilis, because endoscopic and microscopic findings can simulate gastric cancer or lymphoma. Herein are reported two cases of syphilitic gastritis and analysis of the clinicopathological characteristics of 11 cases in Korean subjects, including nine previously reported ones. The two present patients were 25‐ and 32‐year‐old men with epigastric pain. The gastroduodenoscopy features were similar: multiple erosive or ulcerative lesions in whole gastric mucosa. Microscopically, gastric mucosa contained severe chronic active gastritis involving submucosa with crypt abscesses, venulitis and granulomas, suggesting unusual infectious gastritis. Numerous spirochetes were identified on silver staining, and serological tests for syphilis were reactive in both patients. They were treated with benzathin penicillin, and follow‐up gastric biopsy specimens indicated non‐specific chronic gastritis without evidence of syphilis. Of the 11 Korean patients with syphilitic gastritis, six were female and five were male (mean age, 31.7 years). The most common symptom was epigastric pain and all patients had diffuse erosive, ulcerative or infiltrative lesions on gastroduodenoscopy. Syphilitic gastritis should be considered in young patients with epigastric pain who have diffuse gastric involvement such as erosion or shallow ulcer on endoscopy and unusually extensive chronic active gastritis on microscopy, even if the history of syphilis is not clear.     

时间分辨免疫荧光法用于梅毒抗体测定的探讨

目的 通过对时间分辨免疫荧光法（TRFIA）,酶联免疫吸附试验（ELISA）,明胶颗粒凝集试验（TPPA）三种方法测定梅毒特异性抗体（TP）的比较,并对TRFIA法的特异性、灵敏度、线性进行方法学评价,探讨临床常规使用TRFIA方法检测梅毒特异性抗体的可行性.方法 对2012年1月至2012年7月我院256名门诊患者标本分别用TRFIA、ELISA、TPPA三种方法测定TP,并对TRFIA检测系统的灵敏度、批内,批间精密度、线性范围进行测定.结果 TRFIA与TPPA法检测相比其灵敏度为100％,特异性为98.2％.TP阴阳性结果,经配对卡方检验P＞0.05,三种方法无统计学差别.最低检测浓度0.15NCU,相关系数分别为0.95、0.999,高、中、低三种浓度血清的批内、批间精密度CV＜10％.结论 TRFIA法测定TP灵敏度、特异性、方法性能、相关性均好,可作为临床TP常规筛查方法.     

2009年重庆市巴南区美沙酮门诊吸毒人群HIV、HCV和梅毒感染监测分析

目的对重庆市巴南区吸毒人群HIV、HCV和梅毒感染进行监测分析,为制定干预措施提供依据。方法采用酶联免疫吸附试验对抗-HIV、抗-HCV和梅毒进行检测。结果 207名美沙酮替代治疗吸毒人员中,HIV抗体阳性9例,HCV抗体阳性185例,梅毒感染11例。在9例HIV抗体阳性中,男性7例,女性2例;20~40岁之间的6例,40岁以上的3例;采用静脉吸毒的8例,口吸者1例;共用注射器吸毒的4例,未共用注射器吸毒的5例。在185例HCV抗体阳性中,男性153例,女性32例;20~40岁之间的149例,40岁以上的36例;采用静脉吸毒的184例,口吸者1例;共用注射器吸毒的52例,未共用注射器吸毒的133例。在11例梅毒感染中,男性9例,女性2例;20~40岁之间的6例,40岁以上的5例;采用静脉吸毒的6例,口吸者5例;共用注射器吸毒的9例,未共用注射器吸毒的2例。结论 207名美沙酮替代治疗吸毒人员,HIV、HCV抗体阳性和梅毒感染的男性多于女性;20至40岁之间的阳性率高于40岁以上的人员;采用静脉吸毒的阳性率高于口吸人员;共用注射器吸毒的阳性率高于未共用注射器吸毒的人员。     

梅毒螺旋体外膜蛋白Tp0136的制备及在血清学诊断中的应用

目的:为探索梅毒螺旋体(Treponema Pallidum,Tp)外膜蛋白Tp0136在早期梅毒诊断中的价值,利用基因重组技术制备Tp0136蛋白,用于血清抗体的检测。方法:采用PCR扩增Tp0136基因,构建不含信号肽序列的Tp0136基因原核表达载体,SDS-PAGE检测目的蛋白的表达情况,镍离子亲和凝胶分子筛二步法纯化目的蛋白,Western blot检测其免疫反应性,免疫日本大耳白兔评价其免疫原性,免疫双扩检测其效价,以rTp0136为包被抗原的间接ELISA检测早期梅毒血清抗体。结果:重组工程菌包涵体形式表达相对分子质量约为46 KDa的rTp0136,表达率为15%,制备得到纯度大于98%的rTp0136。纯化的rTp0136能诱导大耳白兔产生特异性免疫应答,免疫双扩测得其效价为1:32。Western blot检测重组蛋白能与兔抗Tp0136多克隆抗体发生特异性反应。间接ELISA检测正常人血清均为阴性,而早期梅毒血清抗体的阳性率为75.1%。结论:重组Tp0136具有良好的免疫活性,预示其在早期梅毒血清学诊断中具有良好的前景。