丙型肝炎患者唾液和尿HCV－RNA检测及其意义

采用套式聚合酶链反应检测丙肝患者血清，唾液和尿各５２份。结果血清ＨＣＶ－ＲＮＡ阳性３９例，２３例分别在尿和唾液中检出ＨＣＶ－ＲＮＡ；１３份血清ＨＣＶ－ＲＮＡ阴性者中，尿和唾液均未检出ＨＣＶ－ＲＮＡ，有非常显著差异（Ｐ＝０．０００１）。血清ＨＣＶ－ＲＮＡ阳性者分ＡＬＴ正常组和异常组，尿和唾液ＨＣＶ－ＲＮＡ阳检率分别为１８％和７５％，亦有非常显著差异（Ｐ＝０．００１）。对照组１３例正常人血清、唾液和尿ＨＣＶ－ＲＮＡ均为阴性。表明丙肝患者尿和唾液中能否检出ＨＣＶ－ＲＮＡ与病毒血症及ＡＬＴ有关，提示丙肝患者的尿和唾液可能为ＨＣＶ感染的传播媒介。     

青海省首次发现的2例HIV感染者HIV—1毒株的基因特征和亚型分析

目的：分析青海省首次发现的HIV感染者HIV－1毒株的基因特征及亚型。方法：从2例感染者外周静脉血淋巴细胞中提取前病毒DNA，使用套式PCR方法，扩增HIV的Env基因，对其Env C2-V3及邻近序列进行分析。结果：两个毒株与国际参考序列的Coon间的基因离散率最小为4．93％，而远离其它亚型，2个毒株间的基因离散率为1．35％，进一步的系统树分析显示，2个毒株与Ccon聚集在一起。结论：2个毒株均为C亚型，而且HIV－1毒株刚进入青海省且流行的时间不长。     

Frequency of Hepatitis B Virus infection among Patients before Chemotherapy Treatment

Background: Hepatitis B virus (HBV) is an important public health problem worldwide. Chronic HBV in patients undergoing chemotherapy and immunosuppressive treatment are at risk of HBV reactivation. The consequence of HBV reactivation in immunosuppressed patients may lead to liver failure and death. Therefore, this study was conducted to investigate the frequency of HBV markers in cancer patients before chemotherapy. Materials and Methods: In this study cross-sectional, blood samples were collected from 90 cancer patients before chemotherapy. The patient’s sera were tested for the presence of HBsAg and anti-HBc using enzyme-linked immunosorbent assay (ELISA). The HBVDNA was tested for patient’s sera using nested polymerase chain reaction (nested-PCR). Results: Among 90 patients, 42(46.7%) were males and 48 (53.3%) females, with a mean age of 52.52 ± 11.71 years (range, 25–83 years). Of the 6/90 (6.66%)  patients, including 4/42 (9.5%) males and 2/48 (4.1%) females cases were positive for HBsAg,  anti-HBc and HBV DNA, (P=0.31).  The frequency of HBV infection in cancer patients  was rectal 3(3.33%),  breast cancer  2 (2.22%) and prostate 1(1.11%) cases. The sera of 8/84 (9.52%) patients including 5/39 (12.82%) males and 3/45 (6.66%) females tested positive for anti-HBc, but negative for HBsAg and HBV DNA. (P=0.55). The results of phylogenetic tree revealed that  four isolated HBV DNA in cancer patients were cluster with genotype D. Conclusions: High frequency of 6.66%  HBV infection have been observed in cancer patients before chemotherapy. The sera of  9.52% patients were only positive for anti-HBc IgG which may indicate the past HBV infection or presence of OBI but requires further investigation. To prevent HBV or OBI reactivation, the screening of HBV DNA and anti HBc should be implemented for cancers patients before chemotherapy.     

深圳地区HIV-1母婴垂直传播实验室诊断方法探索及其分子流行病学研究

目的 探索艾滋病病毒Ⅰ型(HIV-1)阳性孕产妇新生婴儿核酸检测的辅助性诊断方法,了解深圳地区母婴传播所感染的HIV-1阳性婴儿HIV-1毒株的分子流行情况,帮助分析HIV-1在该人群中传播的危险因素.方法 收集深圳市2000-2007年19例HIV-1阳性孕产妇的血样,她们所生新生儿中28人的血样,应用套式聚合酶链式反应(Nested-PCR),对该样本膜蛋白基因(env基因)和核心蛋白(gag基因)进行扩增,并对其各基因区核苷酸序列进行测定和分析.结果 28例婴儿中共得到6例PCR诊断为阳性的婴儿样本,6例样本中共存在CRF01_AE和CRF07_BC两种重组毒株以及B一种亚型,其在所有分析样本中的比例分别为4/6、1/6和1/6;在env基因区与对应的流行代表株01_AE.TH.90.CM240的基因离散率为(9.200±1.600)%;CRF01_AE重组株组内离散率为(11.700±4.000)%;在gag基因区与对应的流行代表株01_AE.TH.90.CM240、07_BC.CN.97.97CN001和B.CN.RL42的基因离散率分别为(4.075±0.763)%、(5.500±0.566)%和(7.150±1.485)%;CRF01_AE重组株、CRF07_BC重组株和B亚型组内离散率分别为(4.033±1.692)%、0.800%和1.900%.结论 该方法有望成为HIV母婴垂直传播中新生婴儿的早期辅助性诊断方法.在深圳地区HIV-1阳性婴儿中,HIV-1流行株以CRF01_AE重组亚型为主,其次是B亚型.并且在母婴传播过程中,HIV-1 env和gag基因变异性较小.     

男性性病病人穿通支原体nPCR检测

目的：探讨扬州地区男性性病病人穿通支原体(Mpe)感染状况及Mpe与STD感染的关系。方法：应用套式PCR(nPCR)对扬州地区的76例男性STD者的尿道拭子进行Uu、Mh、Mpn、Mg、Mf、Mpe、Mpi等七种支原体联合检测，并对Mpe阳性标本作了DNA测序分析。结果：76例男性STD患者中有2例Mpe阳性，阳性率为2．6％。其中1例经DNA测序与Mpe(GTU54^T株)标准株完全一致。结论：Mpe是HIV感染的协同因子和AIDS发病的诱发因素，在STD患者尿道拭子标本检出Mpe值得关注。     

宫颈病变女性人类疱疹病毒8型DNA检测结果分析

目的了解宫颈病变女性人类疱疹病毒8型(human herpes virus 8,HHV-8)感染状况,探讨HHV-8感染与宫颈病变的关系。方法随机选择组织学确诊为宫颈上皮内瘤样变(cervical intraepithelial neoplasias,CIN)和宫颈癌的患者310例作为观察组,同区域、同期参加健康体检并排除有宫颈病变的女性310例为对照组,采集血液标本,提取DNA,巢式PCR检测HHV-8 ORF26,核酸电泳观察结果。结果观察组血液样品HHV-8检出率11.8%(33/280),对照组检出率6.9%(20/288),两组差异有统计学意义(P<0.05)。结论宫颈病变女性HHV-8检出率高于正常女性,应加强监测。     

Simultaneous monitoring of CMV and human herpesvirus 6 infections and diseases in liver transplant patients: one-year follow-up

OBJECTIVE:

The aim of this study was to simultaneously monitoring cytomegalovirus and human herpesvirus 6 active infections using nested-polymerase chain reaction and, together with clinical findings, follow the clinical status of patients undergoing liver transplant.

INTRODUCTION:

The human β-herpesviruses, including cytomegalovirus and human herpesvirus 6, are ubiquitous among human populations. Active infections of human herpesvirus 6 and cytomegalovirus are common after liver transplantation, possibly induced and facilitated by allograft rejection and immunosuppressive therapy. Both viruses affect the success of the transplant procedure.

METHODS:

Thirty patients submitted to liver transplant at the Liver Transplant Unit, at the Gastro Center, State University of Campinas, SP, Brazil, were studied prospectively from six months to one year, nested-polymerase chain reaction for cytomegalovirus and human herpesvirus 6 DNA detections. Two or more consecutive positive nested-polymerase chain reaction were considered indicative of active infection.

RESULTS:

Active infection by cytomegalovirus was detected in 13/30 (43.3%) patients, median time to first cytomegalovirus detection was 29 days after transplantation (range: 0-99 days). Active infection by human herpesvirus 6 was detected in 12/30 (40%) patients, median time to first human herpesvirus 6 detection was 23.5 days after transplantation (range: 0-273 days). The time-related appearance of each virus was not statistically different (p = 0.49). Rejection of the transplanted liver was observed in 16.7% (5/30) of the patients. The present analysis showed that human herpesvirus 6 and/or cytomegalovirus active infections were frequent in liver transplant recipients at our center.

CONCLUSIONS:

Few patients remain free of betaherpesviruses after liver transplantation. Most patients presenting active infection with more than one virus were infected sequentially and not concurrently. Nested-polymerase chain reaction can be considered of limited value for clinically monitoring cytomegalovirus and human herpesvirus 6.     

北京地区2006 - 2010年女性HIV-1感染者流行毒株分子特征研究

目的分析北京地区女性HIV-1感染者流行毒株分子特征。方法随机采集2006 - 2010年北京地区新发现女性HIV-1感染者的抗凝全血标本100份,分离血浆,提取病毒RNA,用反转录藻式聚合酶链反应扩增病毒gag基因,并进行序列测定和亚型分析。结果系统进化分析确定北京地区女性HIV-1感染者存在A1(1),A2(1),B(3),B''(23),C(8),G(2),H(1),CRF01_AE(18),CRF02_AG(3),CRF06_cpx(1),CRF07_ BC(4),CRF08_BC(4)和B''/C (3) 13种亚型,其比例分别为1.22%,1.22%, 3.66%, 28.05%, 9.76%, 2.44%,1.22%, 21.95%, 3.66%,1.22%,17.07%, 4.88%和3.66%, B'', CRFO1_AE和CRF07_BC为主要亚型,6种少见亚型(A1,A2,G,H,CRF02_AG和CRF06_cpx)合计占1.0%。结论北京地区女性HIV-1感染者流行毒株遗传多样性高于男性,B’亚型比例高,少见亚型比例也较高。     

Detection of the human Parvovirus B19 in nonimmune hydrops fetalis using immunohistochemistry and nested-PCR in formalin-fixed and paraffin-embedded placenta and fetal tissues

The aim of the present study was to determine whether there is an association between Parvovirus B19 infection and hydrops fetalis setting in fetus and neonate. Twenty-nine samples were analyzed by three methods. Each sample was histologically examined for viral nuclear inclusions in fetal organs and placenta, then immunohistochemical study using Parvovirus B19 antibody that recognized the VP2 protein of the Parvovirus B19 capsid was done in tissue embedded in paraffin (lungs, liver, thymus, kidneys, heart and placenta). Nested-PCR analysis was done after DNA extraction from paraffin blocks and using specific primers of the Parvovirus B19 VP1 gene. Apparent causes of hydrops were eliminated such as metabolic diseases, cardiac failure or malformation. The standard histological study objects viral inclusion in one case (lung tissue). However, the immunohistochemical study was negative in all cases. Nested-PCR demonstrates the presence of the viral DNA in five cases. Our study demonstrates that the implication of Parvovirus B19 in hydrops fetalis must be affirmed by the use of more than one method. Nested-PCR is the most sensitive method in our study and can be easily used for the detection of Parvovirus B19 in formalin-fixed paraffin-embedded tissues.     

孕妇血浆中游离胎儿DNA在产前诊断中的应用

目的探讨孕妇血浆中游离胎儿DNA在非创伤性产前诊断中的可行性。方法用柱分离法提取46例孕妇血浆中的DNA,用巢式PCR技术扩增其胎儿SRY基因,并引入内参照基因ATL1特异序列。结果28例孕男胎的孕妇血浆中有26例经巢式PCR扩增出SRY基因。18例孕女胎的孕妇血浆中有17例经巢式PCR只扩增出ATL1基因,灵敏度和特异度分别为92.9%(26/28)和94.4%(17/18),总符合率93.5%(43/46)。结论应用孕妇血浆中胎儿DNA作产前诊断灵敏度和特异度较高,是一种非创伤性产前诊断方法,具有广泛的临床应用前景。