武强县小学生沙眼流行病学调查

目的调查河北省武强县小学生沙眼的患病率,并分析其危险因素。方法采用随机整群抽样调查方法,抽取武强县7所学校小学生共计1622例,其中男817例,女805例;年龄6～16岁,平均（11.91±2.24）岁。所有被调查对象均进行临床检查,沙眼诊断标准参照世界卫生组织（WTO）制定的沙眼分期标准;从临床诊断为沙眼的患者中,随机抽样进行结膜刮片,镜检法检查沙眼包涵体,利用酶联免疫学与聚合酶链反应（PCR）检测沙眼衣原体;同时选择正常小学生作为正常对照。结果被调查的1622例小学生中,临床诊断为沙眼的379例,患病率为23.4%[95%置信区间（CI）：25.5%～21.3%],其中滤泡性沙眼（TF）307例（81%）,浸润性沙眼（TI）72例（19%）。患病率男为20.9%,女为25.8%,两者比较差异有统计学意义（χ2=5.455,P=0.020）。各年龄组沙眼患病率比较,差异有统计学意义（χ2=9.972,P=0.019）。从379例临床诊断沙眼中随机抽样了168例及正常对照组42例进行结膜刮片检查,沙眼包涵体均为阴性,酶联免疫学检测阳性64例（38.1%）,PCR检测阳性109例（64.9%）。危险因素分析表明沙眼的发生与城郊区居住、性别、年龄有关。结论武强县小学生中仍有沙眼流行,主要以TF为主,需针对危险因素开展沙眼防治。     

大同市小学生沙眼流行病学调查

目的 调查大同市小学生流行性沙眼的患病率,并分析其危险因素.方法 采用随机整群抽样调查方法.抽取大同市小学生共计1236人,其中男生624人,女生612人,年龄6～14岁,平均年龄(10±2)岁.所有被调查对象均进行临床检查,沙眼诊断标准参照世界卫生组织制定的简易沙眼分期标准;对临床诊断为沙眼的患者取结膜标本,进行免疫学与聚合酶链反应(PCR)检测沙眼衣原体.采用χ2检验分析沙眼患病率与学校、性别、年龄的关系,并对实验室检查结果进行统计学分析;采用Logistic回归分析沙眼患病的危险因素,先用Logistic单因素分析相父因素与沙眼患病率之间的关系,以P＜0.05作为入选条件,选取相关自变量,然后应用多因素非条件Logistic回归分析提取主要危险因素.结果 调查的:1236名小学生中,135例临床诊断为沙眼,沙眼患病率为10.9%[95%置信区间(CI):9.2%～12.6%],其中滤泡性沙眼(TF)117例,浸润性沙眼(TI)18例.男生沙眼患病率为8.8%,女生沙眼患病率为13.1%,女生沙眼患病率明显高于男生(P=0.016).各年龄组沙眼患病率比较,差异无统计学意义(P=0.801).临床诊断沙眼的135例小学生中,沙眼农原体免疫学检测阳性13例(9.6%),PcR检测阳性86例(63.7%).危险因素分析表明沙眼的发生与家人是否共用毛巾脸盆及是否经常揉眼睛等因素相关.结论 大同市小学生中仍有沙眼流行,需要针对危险因素开展沙眼防治.     

睑结膜部炎症者肺炎衣原体、沙眼衣原体核酸检测

目的 探讨我国睑结膜部炎症者病灶部是否存在肺炎衣原体（Cpn)。方法 收集39例慢性结膜炎、沙眼患者的睑结膜病灶部刮取物，应用nested PCR(nPCR)技术检测肺炎衣原体（Cpn)和沙眼衣原体（Ct)核酸。并随机抽取2例Cpn nPCR阳性产物进行全自动DNA测序，加以佐证。结果 39例患者中Cpn阳性13例（33．3％），Ct阳性5例（12．8％）。慢性结膜炎和沙眼病例均有Cpn和Ct阳性者。2例Cpn nPCR阳性产物经全自动DNA测序与Cpn(CWL-29株）高度同源。结论 我国睑结膜部炎症者病灶部存在Cpn。     

沙眼患者临床与实验室诊断研究

目的探讨沙眼临床特征，选择适用于临床病因学诊断的实验室检查方法。设计回顾性病例系列。研究对象2003年1月至2006年8月间在北京同仁医院眼科中心临床诊断为沙眼的患者61例。方法沙眼诊断标准参照国内1979年制定的沙眼分期标准，对患者的一般情况、临床表现和实验室检查结果进行了回顾性分析。实验室检查包括结膜刮片查找包涵体、沙眼衣原体抗原检查和PCR检查。主要指标角结膜临床体征，结膜刮片、沙眼衣原体抗原及PCR检查结果。结果临床诊断沙眼患者61例中，男性28例，女性33例，平均年龄（29．05＋19．99）岁；Ⅰ、Ⅱ和Ⅲ期患者分别占88．5％、8．2％和3．3％。刮片检查包涵体阳性检出率11．5％；沙眼衣原体抗原检查阳性检出率为68．9％；PCR检查阳性检出率为78．7％，抗原及PCR检查的检出率高于刮片检出率（P=0．00）。结论门诊沙眼患者主要为Ⅰ期表现，病变较轻，临床需选择敏感的抗原或PCR检查辅助沙眼病因学诊断。     

聚合酶链反应检测泪液中沙眼衣原体的临床研究

目的探讨泪液聚合酶链反应(polymerase chain re-action,PCR)快速检查沙眼衣原体的临床价值。方法清洁结膜囊和睑缘后,微量移液管在内眦角吸取泪液100μL,应用PCR扩增技术检测沙眼衣原体DNA,阳性患者治疗2月后复查。结果30例临床诊断为沙眼患者中24例阳性,阳性患者治疗2月后复查,9例阳性,15例阴性。结论PCR技术检测泪液中沙眼衣原体DNA安全、简便、快速、有较高特异性和敏感性,为临床确诊沙眼和观察疗效提供了客观依据。     

聚合酶链反应对眼分泌物中沙眼衣原体的检测价值

目的：探讨聚合酶链反应在沙眼诊断中的临床应用价值。方法：应用聚合酶反应,引物选用具有衣原体属性的１６ＳｒＲＮＡ基因,扩增检测沙眼衣原体。结果：在９３例诊断为沙眼的患者中,沙眼衣原体阳性率７４．２％;在１７８例诊断非沙眼患者中,沙眼衣原体阳性率３．４％,其检测灵敏度和特异性分别为７４．２％和９６．７％,两组间（沙眼组和非沙眼组）比较有显著性差异（Ｐ〈０．００１）。结论：聚合酶链反应对沙眼分泌物中沙眼     

新生儿沙眼衣原体结膜炎的临床分析

目的 探讨新生儿沙眼衣原体（chlamydia trachomatis,CT)结膜炎的发病情况及临床特点。方法 对117例新生儿结膜炎患者作沙眼衣原体PCR检测，PCR－CR－DNA阳性患儿，作产妇宫颈CT检测。结果 新生儿结膜炎，PCR－CT－DNA阳性率为48．7％，母婴传播率为84．2％，新生儿肺炎并发率36．7％。结论 沙眼衣原体为新生儿结膜炎主要的致病菌，产道感染为CT感染的重要感染途径；沙眼衣原体也为小儿肺炎的主要致病菌。     

我国沙眼防治的启迪与思考

沙眼曾是我国首位的致盲性眼病,经过几代人的不懈努力,终于在今年宣布消灭致盲性沙眼。沙眼衣原体的发现相关研究为防控沙眼提供了科学依据。消灭致盲性沙眼这项成就与政府的高度重视及创新性的在中国成功推广“SAFE战略”密不可分。今后仍应强化公共卫生领域疾病的诊断标准和知识培训,流行病学调查应采用国际标准。虽然中国已消灭了致盲性沙眼,但仍应重视沙眼的全面防控,防止再次流行。（中华眼科杂志,2015,51：484-486）     

陕西省活动性沙眼快速评估调查研究

目的:对陕西省沙眼高发流行疑似区活动性沙眼流行情况进行基线评估,为沙眼的预防和治疗提供依据.方法:采用世界卫生组织沙眼简化分级系统,于2013-10-12/2014-01-26由陕西省防盲办公室组织流调队在榆林佳县坑镇、延安延长、汉中南郑县及商洛市商州区4个沙眼高发流行疑似区,抽取年龄6~8岁小学生共计200人,进行沙眼快速评估调查.结果:受检小学生200人中,未检出活动性沙眼病例.结论:在调查区域,未发现活动性沙眼流行区;活动性沙眼在陕西省内不再属于公共卫生问题.     

衣原体单克隆抗体胶体金法测定沙眼衣原体

目的:介绍一种沙眼衣原体的快速检测方法。方法:采用衣原体单克隆抗体胶体金法进行实验室诊断及鉴别诊断沙眼及其它结角膜病。结果:阴性结果,仅检测板的质控区(C)有1条红线,检测区(T)无红线出现;阳性结果,除质控区(C)外,另有1条红线出现在检测区(T)。10例临床诊断沙眼的患者,其检测结果阳性率为100%。10例其他原因引起的慢性结膜炎患者,其中1例查到沙眼衣原体。10例慢性病毒性结膜炎患者中1例查到沙眼衣原体。结论:该方法简便快速,仅0.5h可出结果,结果判断直观,不需要荧光显微镜、酶联检测仪等设备。具有广阔的推广应用前景。     

Multiplex polymerase chain reaction for diagnosis of viral and chlamydial keratoconjunctivitis

PURPOSE: To develop a multiplex polymerase chain reaction (PCR) for the detection of adenovirus, herpes simplex virus, and Chlamydia trachomatis in conjunctival swabs. METHODS: Oligonucleotide primers for detection of the 3 agents were combined in one reaction and evaluated for optimal performance using control DNAs of adenovirus type 2, herpes simplex virus, and C. trachomatis plasmid. The multiplex PCR was evaluated prospectively against its corresponding uniplex PCRs, virus isolation, Chlamydia Amplicor PCR, and an immunoassay technique (immune dot blot test) in a total of 805 conjunctival swabs from patients with suspected viral and chlamydial keratoconjunctivitis. RESULTS: The multiplex PCR was as sensitive as uniplex PCRs for the detection of the agents in clinical specimens. In the prospective study, 48 of 49 (98%) clinical specimens were positive for adenovirus by the multiplex PCR compared with 26 of 49 (53%) by adenovirus isolation. For herpes simplex virus detection, the multiplex PCR had a sensitivity of 92% (34/37) compared with 94.5% (35/37) by cell culture. The multiplex PCR produced identical results to the Amplicor PCR (21/21; 100%) compared with 71% (15/21) by the immune dot blot test. CONCLUSIONS: With clinical specimens the multiplex PCR was as sensitive as its respective uniplex PCRs but more sensitive than adenovirus isolation and as sensitive as herpes simplex virus isolation or C. trachomatis Amplicor PCR. It has the potential to replace several diagnostic tests with consequent savings in cost. The test also reduces the risk of misdiagnosis by the clinicians.     

Identification of 18S ribosomal DNA genotype of Acanthamoeba from patients with keratitis in North China

PURPOSE: To identify the genotype of 18S ribosomal RNA gene (18S rDNA, Rns) of the Acanthamoeba strains isolated from patients with keratitis in northern China. METHODS: The genus-specific primers JDP1 and JDP2 were used for the amplification of the amplimer ASA.S1. With DNA PCR and sequencing, Rns genotypes were identified according to the DF3 sequence variances. RESULTS: Of all 26 DF3 sequences obtained from the 26 Acanthamoeba strains, 18 were unique (69.2%). Of those, 17 were Rns genotype T4 and one was Rns genotype T3. CONCLUSIONS: Most Acanthamoeba strains isolated from keratitis in northern China were Rns genotype T4. The results are in agreement with recent results in the literature.     

Molecular epidemiology of trachoma in a Gambian village.

The application of a diagnostic and genotyping technique based on the polymerase chain reaction (PCR) to the study of trachoma epidemiology in the Gambian village of Jali is reported. PCR based on the major outer membrane protein (MOMP) gene of Chlamydia trachomatis appears to be more sensitive than either isolation or antigen detection by enzyme immunoassay; it had a specificity of 95% and sensitivity of 51% against clinical signs. PCR genotyping identified genotypes A and B of Chlamydia trachomatis circulating in Jali. Sequencing revealed a Pst1 restriction endonuclease site in the amplified MOMP gene of some B strains but not others; Pst1 digestion of the PCR product proved an easy method of distinguishing these strains. The distribution of serotypes and B strain variants shows a significant degree of household clustering (p < 0.001). PCR based genotyping combined with strain typing provides a new and powerful epidemiological tool for the study of transmission events in trachoma.     

Comparison of an rRNA-based and DNA-based nucleic acid amplification test for the detection of Chlamydia trachomatis in trachoma

BACKGROUND/AIM: The World Health Organisation (WHO) hopes to achieve global elimination of trachoma, still the leading cause of preventable blindness worldwide, in part through mass antibiotic treatment. DNA-based nucleic acid amplification tests (NAATs) are currently used to evaluate the success of treatment programmes by measuring the prevalence of C trachomatis infection. Some believe that newer ribosomal RNA (rRNA)-based tests may be much more sensitive since bacterial rRNA is present in amounts up to 10 000 times that of genomic DNA. Others believe that rRNA-based tests are instead less sensitive but more specific, due to the presence of dead or subviable organisms that the test may not detect. This study compares an rRNA-based test to a DNA-based test for the detection of ocular C trachomatis infection in children living in trachoma-endemic villages. METHODS: An rRNA-based amplification test and DNA-based polymerase chain reaction (PCR) were performed on swab specimens taken from the right upper tarsal conjunctiva of 56 children aged 0-10 years living in two villages in Amhara, Ethiopia. RESULTS: The rRNA-based test detected ocular C trachomatis infection in 35 (63%) subjects compared with 22 (39%) detected by PCR (McNemar's test, p = 0.0002). The rRNA-based test gave positive results for all subjects that were positive by PCR, and also detected infection in 13 (23%) additional subjects. CONCLUSION: The rRNA-based test appears to have significantly greater sensitivity than PCR for the detection of ocular chlamydial infection in children in trachoma-endemic villages. Using the rRNA-based test, we may be able to detect infection that was previously missed with PCR. Past studies using DNA-based tests to assess prevalence of infectious trachoma following antibiotic treatment may have underestimated the true prevalence of infection.     

补体因子H基因单核苷酸多态性与渗出型老年性黄斑变性易感性的相关性研究

目的 探讨渗出型老年性黄斑变性(AMD)易感性与补体因子H(CFH)基因单核苷酸(SNP)多态性的相关性。方法 病例对照研究。136例渗出型AMD患者（AMD组）和年龄、性别与之匹配的140名正常健康者（对照组）纳入研究。取得所有受检者的知情同意后,抽取晨起空腹肘静脉血4 ml,提取基因组DNA。采用多聚酶链反应和特异性限制内切酶消化法检测CFH Y402H(rs1061170)、CFH-257C＞T(rs3753394)及CFH IVS15(rs1329428)的基因型和等位基因。采用SHEsis软件构建单倍型,对比分析两组CFH基因SNP不同单倍型的频率。分析CFH基因SNP不同等位基因、基因型和单倍型与渗出型AMD的相关性。结果 CFH Y402H(rs1061170)存在TT、TC、CC 3种基因型,等位基因位于位点T、C;CFH-257C＞T(rs3753394)存在CC、CT、TT 3种基因型,等位基因位于位点C、T;CFH IVS15(rs1329428)存在AA、AG、GG 3种基因型,等位基因位于位点A、G。AMD组、对照组CFH基因型和等位基因频率比较,差异均有统计学意义(P＜0.05)。CFH Y402H (rs1061170)的杂合子基因型TC、CFH-257C＞T(rs3753394)的纯合子基因型TT及CFH IVS15(rs1329428)的纯合子基因型GG均与渗出型AMD有相关性(OR=4.11,2.55,3.11;P＜0.05);等位基因T、C、G为风险等位基因(OR=3.14,1.72,1.79;P＜0.05)。AMD组和对照组单倍型TCG、CTG及CTA频率间差异有统计学意义(X2=10.53,6.60,32.82;P＜0.05);其余单倍型频率间差异无统计学意义(P＞0.05)。结论 CFH基因SNP多态性与渗出型AMD易感性有关。     

Pesky trachoma suspect finally caught

AIM: Face seeking flies have long been thought to transmit Chlamydia trachomatis, the causative agent of trachoma, but this has never been proven. The four criteria proposed by Barnett, previously used to incriminate other arthropods suspected of transmitting disease, were examined. One of these criteria remains unmet: the repeated demonstration of the presence of C trachomatis on flies. The authors used polymerase chain reaction (PCR) to look for the presence of C trachomatis DNA on flies in the Gurage Zone of Ethiopia. METHODS: Using sticky paper, one fly was collected from the face of each of 103 children aged 1-10 years. The piece of fly paper to which the fly was attached was cut out, followed by the collection of an empty piece from an arbitrary area of the fly paper, which served as control. Roche Amplicor PCR kits were used to detect C trachomatis DNA. RESULTS: Evidence of C trachomatis by PCR was found on 15 of 103 flies versus 0 of 103 controls (p = 0.0001). CONCLUSION: These results meet the final criterion needed to incriminate flies as a vector of trachoma. However, interventional studies will be needed to show the importance of fly control.     

糖尿病视网膜病变黄斑囊样水肿与诱导型一氧化氮合成酶G-954C基因多态性

目的:分析中国人诱导型一氧化氮合成酶G-954C等位基因位点的基因表型在糖尿病视网膜病变黄斑囊样水肿患者与正常对照的差异。方法:病例对照,89例糖尿病视网膜病变黄斑囊样水肿的患者与年龄及性别匹配的90例健康对照纳入该项研究。研究采用了巢式聚合酶链式反应、限制性核酸内切酶技术、基因片段的序列测定。主要检测指标为诱导型一氧化氮合成酶G-954C位点的基因表型。结果:89例患者和90例健康对照的诱导型一氧化氮合成酶G-954C位点的基因表型全部检测结果为GG型。结论:诱导型一氧化氮合成酶G-954C位点的基因多态性可能在中国人为较低的变异频率,而且可能与糖尿病视网膜病变黄斑囊样水肿的易感性关系不大。     

角膜炎棘阿米巴的18S核糖体DNA基因型鉴定

目的 鉴定角膜炎棘阿米巴分离株的18S核糖体DNA(18S rDNA,Rns)基因型。方法 分离转种培养26株致角膜炎的棘阿米巴虫株,提取虫株全基因组DNA,应用棘阿米巴属特异性引物,通过聚合酶链反应(PCR)扩增部分18S rDNA序列。扩增产物测序后,采用分子生物学软件Clustal X和MEGA2对所测序列进行分析,与基因库中已有序列进行比较,分析北京市眼科研究所保存的棘阿米巴虫株Rns基因型特点及与其他国家和地区虫株的基因型异同。结果 分离保存的26株感染角膜的棘阿米巴虫株中,25株(96％)属于Rns T4基因型,1株(4％)属于Rns T3基因型。结论 根据Rns基因特点,感染角膜的棘阿米巴虫株多属于T4基因型,部分虫株属于T3基因型,此分型结果与国外的相似,感染角膜的棘阿米巴可能与特定的基因型有关。(中华眼科杂志,2004．40：389-394)