Optimised sample preparation of synovial fluid for detection of Chlamydia trachomatis DNA by polymerase chain reaction

OBJECTIVE: To optimise sample preparation of synovial fluid for Chlamydia trachomatis (CT) specific polymerase chain reaction (PCR). METHODS: Serial dilutions of purified CT elementary bodies in synovial fluid were prepared. The synovial fluid pellet was processed by eight different methods of sample preparation. Then samples were analysed by CT specific PCR. The sensitivity of PCR was the basis of ranking of the eight different methods. RESULTS: Highest sensitivity was achieved by methods including an additional step of DNA isolation. Additional extraction of protein and polysaccharides by cetyltrimethylammonium bromide (CTAB) increased sensitivity. Addition of hyaluronidase did not increase sensitivity of QIAEX-DNA extraction but was necessary, however, before phenol-chloroform-DNA extraction. CONCLUSIONS: The method of synovial fluid sample preparation significantly influences the sensitivity of subsequent PCR. Additional DNA isolation and extraction of PCR inhibitors by CTAB led to higher sensitivity.     

Risk of coinfection with Chlamydia trachomatis and Neisseria gonorrhoeae in Nova Scotia

BACKGROUND:

The frequency of Chlamydia trachomatis and Neisseria gonorrhoeae coinfection can vary depending on their individual incidence and prevalence rates.

OBJECTIVE:

To determine the frequency of C trachomatis and N gonorrhoeae coinfections by evaluating the results of testing in 2007 and 2008 to better inform testing and treatment decisions.

METHODS:

Specimens from the same patient submitted on the same day served as the basis for the present study. The age, sex and the source of the specimen were also linked to the accession number. Infection and coinfection rates were analyzed in both males and females.

RESULTS:

Concurrent testing was performed on 41,567 female specimens and 1827 male specimens, of which, 1495 female samples (3.6%) tested positive for C trachomatis infection and 88 (0.2%) tested positive for N gonorrhoeae infections. Only 31 females were coinfected; however, for those between 11 and 25 years of age, 25 of 61 females (40.1%) with N gonorrhoeae infection also tested positive for C trachomatis infection; conversely, 25 of 1248 females (2.0%) with C trachomatis infection also tested positive for N gonorrhoeae infection. For males, 213 (11.7%) tested positive for C trachomatis infection, and 59 (3.2%) tested positive for N gonorrhoeae infection. In 30 males with N gonorrhoeae between 11 and 25 years of age, and 149 males with C trachomatis, eight coinfections were observed (26.7% and 5.3%, respectively). Of those older than 25 years of age, only five of 905 men and six of 19,465 women were coinfected. None of the 10,935 women who were 30 years of age or older had coinfections.

CONCLUSION:

The N gonorrhoeae coinfection rate in males with C trachomatis may justify empirical antimicrobials; however, in females, the proportion of coinfected may not justify empirical treatment for N gonorrhoeae infection when the C trachomatis test is positive and N gonorrhoeae testing has not been performed.     

MoPn沙眼衣原体致小鼠生殖道感染模型的初步研究

目的 探讨建立沙眼衣原体(Ct)生殖道感染小鼠模型的接种剂量，观察感染小鼠阴道的排菌规律和小鼠输卵管病交。方法 不同数量的鼠肺炎型沙眼衣原体(MoPn)阴道内接种雌性小鼠，每只小鼠感染后3d及每周取阴道拭子作细胞培养分离病原体，并进行包涵体形成单位(IFU)记数和统计小鼠培养阳性率。观察小鼠输卵管病交并用电镜检测。结果 阴道内接种MoPn可致小鼠感染，取阴道宫颈拭子可分离到Ct，5～6W后停止排菌。实验中发现用10^5 IFU以下Ct阴道内接种，不能使感染组小鼠出现明显发病，而用10^6和10^7 IFU接种小鼠，小鼠出现明显发病，电镜直接观察到Ct存在于输卵管病交组织。结论 阴道接种Ct可逆行感染上生殖道感染，导致小鼠输卵管阻塞和输卵管积水，10^6 IFU是建立上生殖道感染小鼠模型的适合感染IFU。     

广州地区女性不同人群沙眼衣原体感染调查及危险因素分析

目的 调查广州地区不同人群女性泌尿生殖道沙眼衣原体(CT)感染情况,分析CT感染的危险因素.方法 由医护人员组成性病艾滋病高危人群干预队,为目标人群提供性病防治知识讲座、问卷调查,发放健康处方和安全套,采集生殖道拭子用于萘瑟氏淋球菌(GC)、CT、解脲/人型支原体(UU/MH)等的常规检测.结果 共问卷调查788人,对715人采集了宫颈拭子.CT总感染率为10.5%(75/715),其中流动人口育龄妇女、妇科门诊就诊者、暗娼的CT感染率分别为6.4%(15/234),11.1%(31/279),14.4%(29/202),各组间差异有统计学意义(P<0.05).CT合并GC、UU/MH、单纯疱疹病毒Ⅱ型(HSV-2)、粘液脓性宫颈炎(MPC)感染者分别为5.3%、57.3%、36.0%、32.0%.暗娼中CT合并GC、HSV-2、MPC感染者分别为10.3%、51.7%、27.6%.CT感染与20岁以前发生性行为、未婚、配偶从事商业服务业、中重度宫颈炎以及每周性生活频率超过3次等5个因素有关.固定人群因素后发现,首次性生活年龄越早、口服避孕药为危险因素.结论 性工作者是性病行为干预的重点,减少性生活频率,加强青少年禁欲宣传有助于减少CT感染的流行.     

10 676例泌尿生殖道感染门诊患者淋球菌、沙眼衣原体和解脲支原体感染分析

目的探讨2016年7月—2017年3月来上海某医院门诊就诊的10 676例泌尿生殖道感染患者淋球菌(Neisseria gonorrhoeae, NG)、沙眼衣原体(Chlamydia trachomatis, CT)和解脲支原体(Ureaplasma urealyticum, UU)的感染分布情况。方法运用实时荧光定量PCR方法对10 676例泌尿生殖道感染患者分别或同时进行NG、CT和UU测定。结果 NG、CT和UU 3种病原体总检出率分别为1.47%、7.57%、71.27%,其中合并感染756例。检出率以21～30岁年龄段患者最高。在男性患者的样本中,CT的检出率为10.60%,UU的检出率为33.05%,NG的检出率为32.70%。在女性患者中,UU的检出率居首位,为72.16%,CT检出率为7.51%,NG检出率最低为0.89%。结论 UU和NG的检出率具有明显的性别差异。随着患者年龄的增长,3种病原体检出的阳性样本数逐渐下降。     

Ribosomal RNA evidence of ocular Chlamydia trachomatis infection following 3 annual mass azithromycin distributions in communities with highly prevalent trachoma

Twelve trachoma-hyperendemic communities were treated with 3 annual mass azithromycin distributions. Children aged 0-9 years were monitored 1 year following the third treatment. An RNA-based test detected ocular chlamydial infection in more children than did a DNA-based test (6.9% vs 4.2%), and in a larger number of communities (8 vs 7).     

Prevalence of HIV and Chlamydia trachomatis infection in 15--19-year olds in rural Tanzania

OBJECTIVE: To estimate the prevalence of HIV and Chlamydia trachomatis (CT) infections amongst adolescents in rural Mwanza Region, Tanzania and their association with demographic variables. DESIGN: Population-based cross-sectional survey. METHODS: All 15--19-year olds living in households selected by random cluster sampling were invited to participate. After interview, urine was tested for HIV and CT. RESULTS: 9445 15--19-year olds were enrolled. HIV prevalence was 0.6% (95% CI: 0.4--0.8%) in males and 2.4% (95% CI: 2.0--2.8%) in females, and increased steeply with age (trend: P < 0.006 and P < 0.001, respectively). After adjustment for age, risk of HIV infection was significantly associated with female sex (OR=4.3), never having been to primary school in males (OR=2.7), and current symptoms of genital discharge (OR=2.3) or genital ulcer (OR=5.3) in females. The prevalence of CT was 1.0% (95% CI: 0.8--1.4%) in males and 2.4% (95% CI: 2.0-2.9%) in females. After adjustment for age, CT infection was associated with female sex (OR=2.4), reported current symptoms of STD (males OR=2.5, females OR=1.9) and positive leucocyte esterase (LE) test (males OR=3.1, females OR=2.6). Eighty-two percent of males and 79% of females with CT were asymptomatic. There was no association between CT and HIV infection in either sex. CONCLUSIONS: There is a high prevalence of HIV and CT amongst adolescents, especially young women, in this rural population, highlighting the need for effective interventions to improve adolescent reproductive health. The high rates of asymptomatic infection imply that innovative strategies are needed to reach and treat young people with STD.     

High prevalence of Chlamydia trachomatis infection among women attending STD and gynecology clinics in Jiangsu province,China: A cross-sectional survey

Epidemics of Chlamydia trachomatis (CT) infection among women are major global public health concerns. This study examined the prevalence of CT infection and associated factors among women attending sexually transmitted disease (STD) and gynecology clinics in Jiangsu province, China.A cross-sectional survey was conducted among women attending STD and gynecology clinics in the province during 2018 to 2019. Socio-demographic and behavioral information were collected through face-to-face questionnaire survey. Cervical swab specimens were collected to test for CT. Chi square tests were used to compare differences in CT prevalence between subgroups of characteristics. Multivariate logistic regression analysis was used to identify factors associated with CT infection.A total of 2664 participants were enrolled. The prevalence of CT infection was 16.6% (95% confidence interval: 15.0%–18.1%). Of those, CT prevalence among participants from STD clinics (19.4%) and South Jiangsu (18.5%) were higher. Female outpatients who were service personnel (adjusted odds ratio [aOR] = 1.667, P = .004) or farmers (aOR = 1.593, P = .039), lived in South Jiangsu (aOR = 1.796, P = .004), and were from STD clinics (aOR = 1.608, P = .022) were more likely to infect CT.Our study showed a high prevalence of CT infection among women attending STD and gynecology clinics in Jiangsu province, China. CT screening, surveillance and treatment promotion should therefore be of top priority on the CT prevention agenda.     

Diagnostic approaches to cytomegalovirus infection in bone marrow and organ transplantation

Cytomegalovirus (CMV) continues to be a clinical problem, impairing the overall success rate of transplantation, either through direct involvement of a variety of end-organs or by inducing indirect effects such as graft rejection. We review here how the virus manages to evade host immune responses and replicate extensively in allograft recipients. Recent studies show that the quantity of CMV (viral load) is related directly to the development of CMV disease. We review how clinically significant levels of CMV viral load can be defined and summarize the results of studies showing that a high CMV viral load is the major determinant of CMV disease, explaining the previously reported risk factors of pre-transplant serostatus and the post-transplant detection of CMV viremia. Note     

Chlamydia trachomatis in human immunodeficiency virus-infected men treated at a referral hospital for sexually transmitted diseases in the Amazonas,Brazil

ObjectivesThe aim of the present study was to determine the Chlamydia trachomatis prevalence and to identify the demographic, behavioural and clinical factors associated with C. trachomatis in human immunodeficiency virus infected men.StudyThis was a cross-sectional study of C. trachomatis prevalence among human immunodeficiency virus-infected men enrolled at the Outpatient clinic of acquired immunodeficiency syndrome of the Fundação de Medicina Tropical Dr. Heitor Vieira Dourado in Manaus, Amazonas, Brazil. C. trachomatis deoxyribonucleic acid from urethral samples was purified and submitted to real time polymerase chain reaction to identify the presence of C. trachomatis.ResultsA total of 276 human immunodeficiency virus-infected men were included in the study. The prevalence of C. trachomatis infection was 12% (95% confidence interval 8.1%–15.7%). The mean age of the participants was 34.63 (standard deviation 10.80) years. Of the 276 human immunodeficiency virus-infected men, 93 (56.2%) had more than one sexual partner in the past year and 105 (38.0%) reported having their first sexual intercourse under the age of 15 years. Men having sex with men and bisexuals amounted to 61.2% of the studied population. A total of 71.7% had received human immunodeficiency virus diagnosis in the last three years and 55.1% were using antiretroviral therapy. Factors associated with C. trachomatis infection in the logistic model were being single (p < 0.034), men having sex with men (p < 0.021), and having previous sexually transmitted diseases (p < 0.001).ConclusionThe high prevalence of C. trachomatis infection among human immunodeficiency virus-infected men highlights that screening human immunodeficiency virus-infected men for C. trachomatis, especially among men having sex with men, is paramount to control the spread of C. trachomatis infection.     

Low Correlation of Serology with Detection of Chlamydia trachomatis by Ligase Chain Reaction and Antigen EIA

Summary The aim of the present study was to evaluate the diagnostic accuracy of serology by using new assays for the detection of genus and species-specific IgG, IgM, IgA and secretory IgA antibody in female sex workers. Cervical swabs and first void urine (FVU) from 314 female sex workers were submitted to nucleic acid amplification by ligase chain reaction (LCx, Abbott). Concomittantly, blood samples were tested for the presence of IgG, IgM and IgA antibodies using a genus-specific assay (rELISA, Medac) and species-specific test (SeroCT, Orgenics). Chlamydia trachomatis infection was detected in a total of 30 (9.6%) female sex workers by LCR. With rELISA, seroprevalences for IgG, IgM and IgA antibody to Chlamydia were 88.9%, 19.1% and 62.7%, respectively. IgG and IgA antibody prevalences against C. trachomatis (SeroCT) were 65.0% and 23.9%, respectively. In comparison to the positive LCR results obtained from cervical swab and/or FVU, the sensitivity of rELISA for Chlamydia IgG, IgA and IgM detection was 93.9%, 83.3% and 16.7%, respectively. With SeroCT, the sensitivity for C. trachomatis-specific IgG and IgA detection was 86.7% and 33.3%, respectively. The specificities of both serologic tests in comparison to LCR were very slow. C. trachomatis infection of the lower genital tract is very low. According to our results, serologic testing for Chlamydia can exclude active infection of the lower genital tract with a high reliability (≥ 95%). However, detection of C. trachomatis can only be reliably achieved by nucleic acid amplification assays. Received: June 25, 1999 · Revision accepted: January 19, 2000     

原发性肝癌患者术后并发肺部感染的危险因素分析

目的回顾性分析原发性肝癌患者术后导致肺部感染的危险因素,为今后临床治疗上采取相应预防措施提供参考依据。方法选取2011年5月-2015年12月于中国人民解放军第一八○医院肝病科就诊的原发性肝癌患者286例,根据术后是否发生肺部感染分为感染组(n=36)和非感染组(n=250)。对可能引起术后肺部感染的诸多因素进行分析,计量资料组间比较采用t检验,计数资料组间比较采用χ~2检验,Logistic多因素回归分析术后并发肺部感染的危险因素。结果单因素分析显示,两组患者年龄分布、吸烟史、糖尿病、术中输血量、腹腔引流管留置时间比较,差异均有统计学意义(P值均0.05);进一步多因素Logistic回归分析显示,吸烟史、糖尿病、术中输血量和腹腔引流管留置时间为患者术后并发肺部感染的独立危险因素(P值均0.05)。麻醉相关因素中使用一次性器械和围手术期抗菌药的使用均使患者术后肺部感染率下降(P值均0.05)。结论吸烟史、糖尿病、术中输血量、腹腔引流管留置时间、麻醉使用一次性器械和围手术期抗菌药的使用情况均为原发性肝癌患者术后并发肺部感染的危险因素,针对危险因素采取相应的防治措施能够有效预防术后肺部感染的发生。     

冠心病与肺炎衣原体感染的临床研究

目的 测定冠心病患者阿奇霉素治疗前后血肺炎衣原体DNA及特异性免疫球蛋白G（immunoglobulinG,IgG）抗体滴度,探讨肺炎衣原体感染与冠心病的相关性。方法 选择150例冠心病患者（冠心病组）及同期临床可疑冠心病而进行冠状动脉造影正常的55例病例作为对照组,应用巢式聚合酶链反应测定外周血单核细胞中肺炎衣原体DNA,用间接微量免疫荧光法测定IgG抗体滴度。冠心病组采用随机、双盲和安慰剂对照的方法分别给予阿奇霉素及安慰剂治疗6个月,于治疗后1周,1,3,6,12个月复查免疫荧光抗体滴度,研究结束时对阳性病例复查肺炎衣原体DNA。结果 冠心病患者肺炎衣原体DNA阳性49例,阳性率33％,对照组仅1例阳性,阳性率为2％,冠心病组与对照组比较阳性率明显增高。冠心病组中55例无症状型冠心病组阳性16例,阳性率为29％,48例不稳定型心绞痛亚组阳性19例,阳性率为40％,47例急性心肌梗死亚组阳性14例,阳性率为30％,冠心病各亚组间阳性率无差异。治疗后DNA转阴率阿奇霉素组为87％,安慰剂组为59％,治疗组疗效明显优于对照组。治疗前IgG抗体滴度冠心病组较对照组明显增高,治疗后无明显变化。结论 冠心病患者肺炎衣原体DNA阳性率及肺炎衣原体特异性抗体增高,阿奇霉素治疗后DNA转阴,抗体滴度无变化,检测DNA临床价值更大。     

Personal and environmental risk factors for active trachoma in children in Yobe state,north‐eastern Nigeria

Objective To determine the personal and environmental risk factors associated with trachoma in pre‐school children in Yobe state, Nigeria. Methods Total of 639 children aged 1–5 from 27 villages were examined in their households for clinical signs of trachoma. Personal and environmental risk factors were independently determined by an observer while clinical examination was being performed. Results The prevalence of active trachoma was 35.7% (95% CI; 32.0–39.6) with prevalence of disease in boys being similar to girls. The highest prevalence was in children 2–3 years of age. The presence of flies on the face (OR 4.14; CI 2.72–6.29) and lack of a toilet in the compound (OR 2.11; CI 1.38–3.23) were independent risk factors for trachoma. Conclusion The prevalence of trachoma is high. Provision and use of toilets, education on personal and environmental hygiene improvements need to be emphasized as part of the Surgery Antibiotics Facial cleanliness and Environmental improvement strategy.     

Cytokine profile in serum and synovial fluid of arthritis patients with Chlamydia trachomatis infection

Chlamydia trachomatis (Ct)-induced arthritis (CtIA) is characterized by persistent Ct infection, which stimulates secretion of cytokines in vitro. We therefore investigated whether CtIA patients have a unique cytokine profile in synovial fluid or serum in vivo. Because underlying Ct infection is overlooked in a high percentage of patients with initially diagnosed undifferentiated oligoarthritis (UOA), we examined whether determination of cytokines might also be of diagnostic relevance for this arthritis form. Matched serum and synovial fluid specimens from 26 patients with CtIA were analyzed and compared to those from 34 patients with UOA in whom Ct infection was excluded and those of nine patients with rheumatoid arthritis (RA). In 15 CtIA patients, Ct DNA from synovial fluid could be amplified by polymerase chain reaction. The following cytokine or cytokine antagonists were measured by enzyme-linked immunosorbent assay: interleukin (IL)-1, tumor necrosis factor (TNF)-, IL-6, IL-1 receptor antagonist, and soluble TNF receptor p75. No statistically significant differences in cytokine levels between patients with CtIA or the other arthritis forms were detected. Also, comparison between CtIA patients with (n=17) and without Chlamydia DNA (n=9) in synovial fluid revealed no significant differences for these cytokines. Cytokine levels in serum and synovial fluid were not different between CtIA, UOA without Ct infection, and RA patients. The intracellular presence of Ct was not associated with a specific profile of these cytokines in vivo.