HBV基因型分布与慢性乙肝临床关系的研究

目的了解淮北市慢性乙型肝炎患者血清中乙型肝炎病毒（HBV）基因型的分布情况,HBV基因型与临床的关系。方法选择HBV-DNA阳性的慢性乙型肝炎（71例）和肝硬化（22例）患者血清93份,采用巢式PCR法检测HBV基因型。结果本组患者HBV基因分型结果为C型57例（61.3%）,B型16例（17.2%）,B/C混合型20例（21.5%）;C、B和B/C基因型HBV分布在性别上无统计学差异;在慢性乙型肝炎和肝硬化两组中C型比例分别为66.2%,45.5%,差异无有统计学意义（P〉0.05）;C、B和B/C基因型患者HBeAg阳性率分别为79.0%（45/57）,68.8%（11/16）,65.0%（13/20）,差异均无统计学意义（P〉0.05）。B/C基因型与B型和C型基因患者在HBV-DNA和ALT水平上有显著统计学意义（P〈0.01）。结论淮北地区HBV优势基因型以C型为主,B/C混合型次之,有少量的B基因型,B/C混合基因型HBV在病毒复制和肝脏损害方面较B型和C型严重。     

Failure to demonstrate human papillomavirus DNA in epithelial ovarian cancer by general primer PCR

One of the recent controversies with substantial clinical interest is the role of HPV in pathogenesis of ovarian cancer. The available highly conflicting data are based on analysis of 175 ovarian carcinomas so far. As an attempt to further elucidate this issue, the first systematic study of HPV detection in ovarian cancer was carried out using a highly sensitive general primer PCR (confirmed by hybridization for low- and high-risk HPV types separately) in a series of 98 histologically and clinically well-characterized epithelial ovarian malignancies. Despite the high (fg) sensitivity and a wide HPV type coverage of the technique used, all 98 ovarian carcinomas failed to demonstrate any signs of HPV DNA whatsoever. The preexisting 12 reports comprising a total of 175 ovarian tumors analyzed for HPV were summarized, giving highly discrepant results (i.e., detection rates from 0 to 100%) with the overall HPV DNA detection rate of 25.7%. The reasons for these discrepant findings are most probably technical. Our data are consistent with those of the majority of the most recent reports failing to disclose HPV DNA in ovarian neoplasia. The present completely negative results make the authors inclined to conclude that HPV is highly unlikely to play any causal role in the pathogenesis of epithelial ovarian neoplasia.     

Novel gamma-2-herpesvirus of the Rhadinovirus 2 lineage in gibbons

We obtained 475 nucleotides of the DNA polymerase gene of a novel human herpesvirus 8 homolog sequence in a gibbon. The finding of this new gibbon virus, which clusters with a related chimpanzee virus in the rhadinovirus 2 genogroup, suggests the existence of a novel gamma-2-herpesvirus in humans.     

用TaqMan探针实时定量PCR法快速检测鼠疫耶尔森菌

目的建立一种快速、准确、特异的定量检测鼠疫耶尔森菌的方法。方法针对鼠疫耶尔森菌特异的染色体标识序列设计引物和TaqMan探针，进行实时定量聚合酶链反应(polymerase chain reaction，PCR)分析，优化反应体系，检测其灵敏度、特异性和重复性。结果以EV76灭活培养物为对象，检测灵敏度可达每反应体系0．08CfU；30株非鼠疫菌株的检测结果表明，对照菌株均为阴性，检测特异性良好；对同一样品进行17次重复检测，其循环阈值的标准偏差为0．35。结论TaqMan探针法能够灵敏、特异、稳定地对鼠疫耶尔森氏菌进行定量检测，为鼠疫监控、诊断提供有力手段。     

格林-巴利综合征与人类白细胞抗原关联的研究

目的 研究格林－巴利综合征（GBS）两种不同亚型（AIDP和AMAN）与HLA－Ⅰ类和Ⅱ类等位基因分型的关系,探讨AIDP和AMAN患者免疫遗传的特点。方法 用改良快速盐析法抽提基因组DNA,采用聚合酶链反应－顺序特异性引物法（PCR－SSP）对64例AIDP、AMAN患者和132例健康人进行HLA－Ⅰ类和Ⅱ类等位基因分型。结果 AIDP组HLA－Ⅰ类基因中A33的频率较对照组升高,相对风险率（RR）为6．13,校正的P值（Pc)为0.011(P值＜0.05)。HLA－Ⅱ类基因中,DR16和DQ5的频率较对照组升高,RR值分别为8.28和3.47,Pc分别为0.014和0.025（P值均＜0.05);AMAN组HLA－Ⅰ类基因中B15、B35频率较对照组升高,RR值分别为4.09和7.08,Pc分别为0.015和0.0008(P值均＜0.05)。结论 HLA－A33、DR16和DQ5与AIDP的易感性可能有关。HLA－B15、B35与AMAN的易感性可能有关。未发现HLA－Ⅱ类基因与AMAN有明显的关联。     

Molecular heterogeneity of glucose-6-phosphate dehydrogenase deficiency in Malays in Malaysia

Multiplex polymerase chain reaction (PCR) using multiple tandem forward primers and a common reverse primer (MPTP) was recently established as a comprehensive screening method for mutations in X-linked recessive diseases. In the work reported here, MPTP was used to scan for mutations of the glucose-6-phosphate dehydrogenase (G6PD) gene. Mutations in exons 3,4,5,6,7,9, 11, and 12 of the G6PD gene were screened by MPTP in 93 unrelated Malaysian patients with G6PD deficiency. Of the 93 patients, 80 (86%) had identified mutations. Although all of these were missense mutations, identified nucleotide changes were heterogeneous, with 9 mutations involving various parts of the exons. These 9 mutations were G-to-A nucleotide changes at nucleotide 871 of the G6PD gene (G871A), corresponding to G6PD Viangchan, G6PD Mediterranean (C563T), G6PD Vanua Lava (T383C), G6PD Coimbra (C592T), G6PD Kaiping (G1388A), G6PD Orissa (C131G), G6PD Mahidol (G487A), G6PD Canton (G1376T), and G6PD Chatham (G1003A). Our results document heterogeneous mutations of the G6PD gene in the Malaysian population.     

Detection and identification of Toscana and other phleboviruses by RT-nested-PCR assays with degenerated primers

Phleboviruses are a large and widespread group of viruses that are transmitted by arthropods. Toscana virus is one of the principal agents that causes meningitis in humans during the summer in Italy and, possibly, in other Mediterranean countries. Rift Valley Fever virus can cause serious illness in both animals and humans, leading to high morbidity and mortality, and is considered to be a potential agent for epizootics and human epidemics. Since information on this group of viruses is still scant, reliable laboratory tools for diagnosis and epidemiological surveillance must be developed, in order to ascertain their real impact on Public Health. Sequence data obtained from Spanish isolates of Toscana virus and other phleboviruses confirmed that natural genome variability may hamper the diagnosis of these agents by molecular methods, so this must be borne in mind when developing reliable assays. In view of the above, a novel and useful protocol has been developed for the detection and specific identification of every member of the phlebovirus genus present in a sample, including Toscana virus, based on a generic RT-nested-PCR, followed by sequencing of the amplified fragment. A change in this method also allowed specific direct detection and identification of wild isolates of Toscana virus of different geographical origin, using newly designed primers. Testing clinical samples with these assays confirmed the role of Toscana virus as an agent that causes acute aseptic meningitis in the central region of Spain.     

Eleven human platelet systems studied in the Vietnamese and Ma'ohis Polynesian populations

The frequencies of human platelet antigens (HPA) are variable among different ethnic groups. Platelet phenotyping and genotyping in different populations are important to the clinical implications of antiplatelet alloimmunization. No report on HPA prevalence has been published concerning the Vietnamese Kinh and Ma'ohis Polynesian populations. Recent anthropological and genetic marker studies suggest that these two groups have a common origin in East Asia, so we have conducted a combined study concerning the frequency of HPA-1 to HPA-11w systems (excluding HPA-8w) and Gov in these two populations. The results demonstrate a similar pattern of prevalence between Ma'ohis and most of the Asian populations. However, it should be noted that the frequency of HPA-2 is closer to northern Caucasian frequencies than to Asian frequencies. The population of Kinh shows an HPA distribution that is closer to the Chinese population than to the northeastern Thais except for HPA-3, closer to the Indonesian population. Given HPA-3 gene frequency distribution fetomaternal incompatibility could occur more frequently with the risk of alloantibody production.     

Polymorphism and distribution of HLA-DR2 alleles and haplotypes in a Greek population

Abstract: HLA-DR2 serological subtyping has indicated that the DR16 serotype appears at a higher frequency relative to the DR15 serotype in the Greek population, differing from the distribution observed in most other Caucasian groups. In this study, we have analyzed by the PCR-SSP technique a DR2-positive group of unrelated Greek individuals selected from our normal control panel for the different DRB1, DRB5, DQB1 and DQA1 DR2-associated alleles present. Six of the 50 individuals analyzed were homo-zygous for DR2, contributing a total of 56 haplotypes for DR2. The observed frequencies of the DR2-related DRB1 alleles were as follows: 58.9% for the DRB1*1601, 7.1% for the DRB1*1602, 25.0% for the DRBl*1501 and 7.1% for the DRB1*1502 allele. The rare allele DRB1*1605 was detected in one heterozygous sample and its presence was definitively established by DNA sequencing. The alleles *1503, *1504, *1505, *1603 and *1604 were not detected. Three DRB5 alleles were identified: DRB5*0202 (67.8%), DRB5*0101 (25.0%) and DRB5*0102 (7.1%). Ten different DRB1/DQB1/ DQA1 DR2-associated haplotypes were denned. The most frequently observed haplotype was DRBl*1601-DQBl*0502-DQAl*0102 (relative frequency =57%) followed by DRB1*1501-DQB1*0602-DQA1*0102 (relative fre-quency=14.3%). In conclusion, the refined analysis of the DR2-associated DRB1 alleles in the Greek population revealed the prevalence of the DRB1*1601 allele. The rare allele DRB1*1605 was demonstrated once. A considerable variety of different DR2-related DR/DQ haplotypes was detected and the overall haplotypic frequencies in the Greek population are distributed differently compared to those reported for most other Caucasian populations.     

Use of nested PCR for differential diagnosis of falciparum malaria reinfection and relapse in drug-resistant patients

Microscopic examination does not allow differentiation of drug-resistant P. falciparum infection relapse from reinfection. However, this differential diagnosis is essential for adequate therapy. Three highly polymorphic P. falciparum genes (msp1, msp2, and glurp) and their alleles reflecting the structural state of these genes were used as genetic markers for differential diagnosis by PCR with internal primers. In 27 patients the characteristics of these alleles were identical before treatment with artersunate and during repeated manifestation of symptoms 14-28 days after the end of therapy, which attested to malaria relapses. In 24 patients the structure of these allele before mefloquine therapy and during repeated manifestation of the symptoms after 2-3 months was different, which attested to reinfection.