成人跖疣人乳头瘤病毒分型及与临床特征的关系

目的 检测成人跖疣乳头瘤病毒(HPV)型别,并探讨HPV分型与临床特征的关系.方法 采用PCR扩增、基因测序和TA克隆技术检测221例跖疣患者疣体的HPV型,记录患者的年龄、病程、症状和疣体数.结果 221例标本中,HPV阳性率为97％(215/221).单一HPV型占96％(206/215),HPV 27、2、57和7(Alpha属)分别占44.7％ (92/206)、12.1％(25/206)、7.3％(15/206)和0.5％(1/206);HPV 65(Gamma属)占0.5％(1/206);HPV 1、63(Mu属)占33.5％ (69/206)和1.5％ (3/206).HPV 27、2和57三型感染者之间年龄、病程、疣体个数、疼痛发生率和性别比较,差异均无统计学意义.与HPV 27感染者相比,HPV 1感染者与年龄≤30岁、病程≤1年、疣体数≤2个和疼痛高发生率相关.结论 HPV 27、2、57和1是感染成人跖疣患者的主要HPV型.HPV型与患者的临床特征间存在相关性.     

子宫颈细胞学检查未见异常的HR-HPV亚型感染妇女的CIN2,3风险分析

目的:探讨子宫颈细胞学检查未见异常的HPV高危亚型感染者的管理模式。方法:收集2010年1月至2012年12月在北京大学第一医院妇产科门诊同时行宫颈细胞学检查及HPV DNA分型检测的妇女的资料,分析初次检出细胞学未见异常者的HPV高危亚型16、18、31、33感染者,其检出CIN2及以上病变的风险以及与感染亚型的相关性。结果:993例细胞学检查未见异常但HPV16、18、31、33型阳性者中,共检出CIN1 76例(7.7%),CIN2 50例(50/993,5.0%),CIN3 27例(27/993,2.7%);其中HPV16(+)感染者532例(532/993,53.6%),检出CIN2 34例(34/532,6.4%),CIN3 21例(21/532,3.9%);HPV18(+)HPV16(-)感染者142例(142/993,14.3%),检出CIN2 2例(2/142,1.4%),CIN3 1例(1/142,0.7%);HPV31(+)HPV16\18(-)感染者137例(137/993,13.8%),检出CIN2 9例(9/137,6.6%),CIN3 2例(2/137,1.5%);HPV33(+)HPV16\18(-)感染者182例(182/993,18.3%),检出CIN2 5例(5/182,2.7%),CIN3 3例(3/182,1.6%)。按是否检出CIN2+进行Logistic回归分析,发现HPV16型感染与CIN2+有相关性[OR值=2.353(95%CI 1.004~5.516),P=0.049]。结论:对筛查中初次检出宫颈细胞学未见异常,但HPV高危亚型感染者应予以重视,对于HPV16、18型感染者建议立即行阴道镜检查。     

耐甲氧西林金黄色葡萄球菌基因多态性分型研究

目的研究耐甲氧西林金黄色葡萄球菌（MRSA）的基因多态性分型特征，为控制感染提供分子流行病学依据。方法利用一条10bp随机引物，建立随机引物多态性（RAPD）分析方法，并利用这一方法对44株MRSA进行基因分型的研究。结果44株MRSA中有30株产生RAPD指纹图谱，经电泳得到2．8条片段，可分为5个型，其中Ⅲ型菌株占50％。结论通过RAPD分型研究，可了解MRSA的基因型流行特征，为控制感染提供分子流行病学依据。     

丙型肝炎病毒逆向点杂交基因分型方法的建立及初步应用

目的 利用逆向点杂交技术建立丙型肝炎病毒(HCV)基因分型新方法。方法在HCV5’端非编码区(5’UTR)设计引物与分型探针，将活性氨基标记的分型探针依次固定在尼龙膜上，制成HCV基因分型逆向点杂交检测膜条。分离纯化血清中的HCV RNA，经逆转录反应和生物素标记引物聚合酶链反应(PCR)巢式扩增后，将扩增产物与检测膜条杂交，过氧化物酶和四甲基联苯胺显色，判断基因分型结果。最后，通过与基因测序结果比较确定新方法的有效性。从佛山地区丙型肝炎患者中随机抽取60份经荧光定量PCR方法对HCV RNA进行过定量分析的血清，用新建方法进行HCV基因分型检测。结果新建HCV逆向点杂交基因分型方法可对所有60份HCV RNA拷贝数在10。～10。／ml之间的抽检血清进行基因分型，发现1b型50例，占83．3％；1a型2例，占3．3％；2a型2例，占3．3％；1a、1b混合型5例，占8．0％；1b、2a混合型1例，占1．7％；未发现2b、3a和3b型。新方法基因分型结果与测序分型结果一致。结论PCR一逆向点杂交技术可以准确有效和简便经济地进行HCV基因分型．适用于临床检测与流行病学研究。在佛山地区人群中感染的HCV以1b型为主。     

广州地区革兰阴性杆菌CTX-M和OXA型广谱β-内酰胺酶基因分型研究

目的调查广州地区临床分离的革兰阴性杆菌中CTX-M型超广谱β-内酰胺酶(ESBLs)和OXA型广谱β-内酰胺酶(beta-lactamase,Bla)的主要基因型别及其流行情况.方法按照NCCLS 2001年标准筛选广州地区临床分离菌株的ESBLs表型;用聚合酶链反应(PCR)扩增法和DNA测序法进行ESBLs基因序列分析.结果PCR扩增结果显示,CTX-M1、CTX-M2、CTX-M9群和OXA的总阳性率在本地区临床分离的临床检测ESBLs阳性的革兰阴性杆菌中分别为9.96%、0、35.5%和1.6%,同时检出≥两种ESBLs基因的菌株数64株,阳性率为5.9%;序列分析进一步证实了CTX-M型ESBLs的具体型别,包括CTX-M-3、CTX-M-22、CTX-M-9、CTX-M-14、CTX-M-17、CTX-M-18、CTX-M-21、CTX-M-24、TOHO-2和TOHO-3,其比例分别为3.23%、3.7%、4.99%、3.32%、2.21%、3.69%、2.95%、4.43%、1.85%和1.48%;其中以CTX-M-9和CTX-M-24阳性率较高;在本地区只检出1种OXA型Bla-OXA-2/PSE-2(1.38%,15/1084);另外,还检出了8株无法具体归类的CTX-M-9型ESBLs,占0.74%;CTX-M型基因主要分布于大肠埃希菌和肺炎克雷伯菌中,分别占42.8%和36.3%,而OXA基因则主要分布于铜绿假单胞菌中,占80%.结论本地区CTX-M类ESBLs也较为常见,其中尤以CTX-M-9和CTX-M-24为主,暂无CTX-M2群ESBLs,可能存在1种或多种新的CTX-M型ESBLs.     

快速盐析法提取DNA在HLA基因分型中的应用

采用快速盐析方法提取人类有核细胞ＤＳＡ，并在临床肾移植的ＨＬＡ基因配型中应用。通过对２１０份不同组织来源的样本与标准酚氯仿法同步对比研究显示：快速盐析法提取模板ＤＮＡ，无污染和有害作用；操作简单、快速，总耗时１１０分钟；方法稳定可靠，无一例失败；所获ＤＮＡ质量好，纯度高，达到酚氯仿法所获ＤＮＡ的水平；用于临床肾移植基因型均获成功，与酚氯仿法结果完全相同。证实该法提取的ＤＮＡ适用于临床器官移植基因水     

Establishment of an optimized set of 406 microsatellite markers covering the whole genome for the Japanese population

Microsatellites, an essential tool for genetic linkage analyses, are selected in genetic studies on the basis of both informativeness and their positions with respect to one another on the genetic map. In order to establish a microsatellite marker set useful for linkage studies in the Japanese population, we first genotyped 64 unrelated Japanese subjects, using 400 microsatellite markers from a commercially available set (ABI PRISM Linkage Mapping Set-MD10) and then determined the allelic frequencies and heterozygosities for these marker loci in the population. In order to optimize the set, we replaced 41 markers having a heterozygosity lower than 0.6 with as many informative markers in the corresponding loci, and newly added six markers in the set to minimize the several gaps found at intervals of over 20 cM. We finally established a set comprising 406 microsatellites with average intervals of 9 cM (maximum, 17 cM) and minimum heterozygosities of over 0.6 (mean, 0.76). All data generated in this study, including the specific polymerase chain reaction (PCR) primer sequences of the newly added markers, are freely available to all researchers at our web site. The genetic tool established here should facilitate genetic linkage studies of various hereditary diseases, especially in the Japanese. Received: December 13, 2000 / Accepted: January 15, 2001     

Genotyping of Hepatitis C virus (HCV) in infected patients from South India

Hepatitis C virus shows substantial nucleotide sequence diversity distributed throughout the viral genome. In the present study genotyping for Hepatitis C virus (HCV) infected patients was based on RFLP analysis of 5' UTR and using type specific primers of NS5B regions. It was observed that 60% of the patients (30 patients with chronic hepatitis) were infected with variants of genotype 1 and 40% of the patients (4 chronic hepatitis patients, 12 patients with chronic renal failure and 4 cirrhosis) were infected with variants of type 3 of HCV. None of the cirrhotic patients and patients with chronic renal failure, in the present study, were infected with type 1 of HCV. While PCR-RFLP, typing was rapid in conjunction with the primers used for RT-PCR, NS5 typing was helpful in determining the subtype. There was good correlation between the two typing methods and this method can be used as a cost-effective method for studying large number of samples. The study shows that predominant genotypes of HCV in South India include type 1 and 3. Type 3 seems to be transmitted nosocomially as suggested by the results in patients with chronic renal failure, as these patients are exposed to multiple medical interventions.     

子宫颈刮片中人乳头瘤病毒的基因分型

目的 确定不同型人乳头瘤病毒（ＨＰＶ）感染的自然历程以及其持续感染在子宫颈癌发展过程中的作用。方法 应用聚合酶链反应检测荷兰１５５名妇女子宫刮颈片中的ＨＰＶ ＤＮＡ，应用线样探针分析法（ＬｉＰＡ）进行，包括ＨＰＶ６，１１，１６，１８，３１，３３，３５，４０，４２，４３，４４，４５，５１，５２，５６和５８的基因分型。结果 １５５例妇女子宫颈片中ＨＰＶ ＤＮＡ检出率为６０％，其中在宫颈细胞学检查正常或     

Role of clinicians and public health professionals for measles surveillance in Canada

The current strategy to eliminate measles in Canada requires stringent surveillance efforts including the laboratory confirmation of measles cases. Clinicians and public health professionals are involved with the events and actions surrounding the initial contact with suspected measles cases, identification of suspected measles cases, collection of the appropriate clinical information, collection of samples for laboratory confirmation and notification of the appropriate public health authority. Clinical information is necessary for the interpretation of laboratory results and is used, together with the laboratory results, to confirm measles cases. Important clinical information required includes the date of onset of symptoms, date of sample collection and measles vaccination history. A blood sample and specimen for virus isolation (urine or nasopharyngeal or throat swab) should be collected within several days of the onset of the rash. In the laboratory, blood samples are necessary for serological confirmation of measles infection, and specimens for virus isolation are necessary for molecular epidemiological purposes.