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A Study of the Technique of Western Blot for Diagnosis of Lyme Disease caused by Borrelia afzelii in China

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Biomedical and Environmental Sciences 2013年第3期26卷 190-200页

作者： LIU Zhi Yun HAO Qin HOU Xue Xia JIANG yi GENG Zhen wu yi mou WAN Kang Lin State Key Laboratory for Infectious Disease Prevention and Control National Institute for Communicable Disease Control and PreventionChinese Center for Disease Control and Prevention Institute of Pathogenic Biology University of South China YuShui District Center for Disease Control and Prevention

Objective To study the technique of Western blot for the diagnosis of Lyme disease caused by Borrelia afzelii in China and to establish the standard criteria by operational procedure. Methods FP1, which is the representative strain of B. afzelii in China, was analyzed by SDS-PAGE, electro transfer and irnmunoblotting assays. The molecular weights of the protein bands of FP1 were analyzed by Gel-Pro analysis software. In a study using 451 serum samples （159 patients with Lyme disease and 292 controls）, all observed bands were recorded. The accuracy of the WB as a diagnostic test was established by using the ROC curve and Youden index. Results Criteria for a positive diagnosis of Lyme disease were established as at least one band of P83/100, P58, P39, OspB, OspA, P30, P28, OspC, P17, and P14 in the IgG test and at least one band of P83/100, P58, P39, OspA, P30, P28, OspC, P17, and P41 in the IgM test. For IgG criteria, the sensitivity, specificity and Youden index were 69.8%, 98.3%, and 0.681, respectively; for IgM criteria, the sensitivity, specificity and Youden index were 47%, 94.2%, and 0.412, respectively.

关键词： Lyme disease Western blot Diagnostic method Borrelia afzelii

Development of ELISAs for the Detection of Urogenital Chlamydia trachomatis Infection Targeting the pORF5 Protein

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Biomedical and Environmental Sciences 2013年第3期26卷 169-175页

作者： LI Zhong Yu HUANG Qiu Lin SU Sheng Mei ZHONG Guang Ming wu yi mou Department of Microbiology and Immunology School of MedicineUniversity of South China Department of General Surgery First Affiliated Hospital of the University of South China Department of Microbiology and Immunology University of Texas Health Science Center at San Antonio

Objective To prepare antibodies against pORF5 plasmid protein of Chlamydia trachomatis develop double-antibody sandwich enzyme-linked immunosorbent assays （DAS-ELISAs） for detection of genital C. trachomatis infections. and the Methods The pORF5 protein was expressed in Escherichia coil and used to immunize BALB/c mice and New Zealand rabbits to produce monoclonal antibodies （mAbs） and polyclonal antibody （pAb） for DAS-ELISAs. Clinical samples from 186 urogenital infection patients （groups I） and 62 healthy donors （groups II） were detected in parallel by the DAS-ELISAs developed in this study and by IDEIA PCE commercial ELISA. Results Two hybridoma cell lines, named 2H4 and 4E6, stably secreting specific mAbs against pORF5 were obtained. The mAb 2H4 was recognized by 32 （17.20%, positive recognition rate） and 25 （13.44%）, mAb 2H4 by 0 （0%） and 2 （3.22%） samples from groups I and II, respectively. The sensitivities of mAbs 2H4 and 4E6 were 92.11% and 77.78% and the specificities were 100% and 96.88%, respectively in relation to the IDEIA PCE commercial ELISA. The sensitivities of detection for the DAS-ELISAs were 10 ng/mL （based on 2H4） and 18 ng/mL （based on 4E6）. Conclusion Two DAS-ELISAs were developed in this study that provided a feasible and effective assay that could be considered alternative tools for the serodiagnosis of C. trachomatis infection.

关键词： Chlamydio trachomatis Monoclonal antibody Polyclonal antibody pORF5 DAS-ELISA

Development and Evaluation of a MAb-Based ELISA for Detection of Chlamy- dophila pneumoniae Infection with Variable Domain 2 and 3 of the Major Outer Membrane protein

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Biomedical and Environmental Sciences 2012年第6期25卷 690-696页

作者： ZHOU Zhou wu yi mou CHEN Li Li LIU Guang Chao LIU Liang Zhuan ZHOU An Wen ZHANG Jun Hua Department of Microbiology and Immunology University of South China Hengyang 421001 Hunan China

Objective This paper aims to develop a monoclonal antibodies （MAbs）- based ELISA for detecting Chlamydophila pneumoniae （C. pneumonioe） antigens in humans with the variable domains （VD） 2 and 3 of the major outer membrane protein （MOMPvD2-VD~） and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae. Methods MOMPvo2-vo3were overexpressed in Escherichia coil and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I （156 patients with typical respiratory illness clinically confirmed before） and Group II （57 healthy donors）. Results In Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II ＂healthy donors＂, 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively. Conclusion The novel ***3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.

关键词： Chlamydophila pneumoniae Major outer membrane protein Monoclonal antibody ELISA

Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization

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Biomedical and Environmental Sciences 2015年第1期28卷 25-35页

作者： GUO Qian YU Yan ZHU Yan Ling ZHAO Xiu Qin LIU Zhi Guang ZHANG Yuan Yuan LI Gui Lian WEI Jian Hao wu yi mou WAN Kang Lin Pathogenic Biology Institute University of South China State Key Laboratory for Infectious Disease Prevention and Control National Institute for Communicable Disease Control and Prevention Chinese Center for Disease Control and Prevention Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases Dalian Maternity and Child Health Care Hospital (the Obstetrics and Gynecology Hospital of Dalian) Key Lab of Laboratory Medicine Wenzhou Medical College

Objective A PCR-reverse dot blot hybridization （RDBH） assay was developed for rapid detection of rpoB gene mutations in ＇hot mutation region＇ of Mycobacterium tuberculosis （M. tuberculosis）. Methods 12 oligonucleotide probes based on the wild-type and mutant genotype rpoB sequences of M. tuberculosis were designed to screen the most frequent wild-type and mutant genotypes for diagnosing RIF resistance. 300 M. tuberculosis clinical isolates were detected by RDBH, conventional drug-susceptibility testing （DST） and DNA sequencing to evaluate the RDBH assay. Results The sensitivity and specificity of the RDBH assay were 91.2% （165/181） and 98.3% （117/119）, respectively, as compared to DST. When compared with DNA sequencing, the accuracy, positive predictive value （PPV） and negative predictive value （NPV） of the RDBH assay were 97.7% （293/300）, 98.2% （164/167）, and 97.0% （129/133）, respectively. Furthermore, the results indicated that the most common mutations were in codons 531 （48.6%）, 526 （25.4%）, 516 （8.8%）, and 511 （6.6%）, and the combinative mutation rate was 15 （8.3%）. One and two strains of insertion and deletion were found among all strains, respectively. Conclusion Our findings demonstrate that the RDBH assay is a rapid, simple and sensitive method for diagnosing RIF-resistant tuberculosis.

关键词： Mycobacterium tuberculosis Rifampin-resistance Reverse dot blot hybridization

肺炎衣原体与中枢神经系统疾病相关性研究进展

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中国人兽共患病学报 2021年第4期37卷 368-372,377页

作者：袁帅吴移谋南华大学病原生物学研究所衡阳421001

肺炎衣原体(Chlamydia pneumoniae,Cpn)是一类具有独特双相发育周期的专性胞内寄生菌。Cpn感染过程复杂且致病类型广,除引起常见的呼吸道感染外,还可引起多个系统的慢性持续性感染。有文献报道,Cpn与中枢神经系统疾病的发生、发展密切... 详细信息

肺炎衣原体(Chlamydia pneumoniae,Cpn)是一类具有独特双相发育周期的专性胞内寄生菌。Cpn感染过程复杂且致病类型广,除引起常见的呼吸道感染外,还可引起多个系统的慢性持续性感染。有文献报道,Cpn与中枢神经系统疾病的发生、发展密切相关。本文对Cpn与中枢神经系统疾病的相关性研究进展进行综述,为今后更合理地指导临床治疗提供重要信息。

关键词：肺炎衣原体中枢神经系统多发性硬化阿尔兹海默病动脉粥样硬化性血管疾病

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棘阿米巴副衣原体病原学研究进展

中国人兽共患病学报 2012年第11期28卷 1137-1142页

作者：刘彦吴移谋南华大学寄生虫学教研室衡阳421001 南华大学微生物及免疫学教研室衡阳421001

目的棘阿米巴副衣原体是1999年被正式命名的新型衣原体,目前已发现8个种的棘阿米巴副衣原体。棘阿米巴副衣原体为棘阿米巴内共生体,复制周期包括原体、网状体和新月体3个阶段,可进行分离培养。棘阿米巴副衣原体病原学的研究为临床、检... 详细信息

目的棘阿米巴副衣原体是1999年被正式命名的新型衣原体,目前已发现8个种的棘阿米巴副衣原体。棘阿米巴副衣原体为棘阿米巴内共生体,复制周期包括原体、网状体和新月体3个阶段,可进行分离培养。棘阿米巴副衣原体病原学的研究为临床、检测和防治提供了重要线索和途径。本文就棘阿米巴副衣原体病原学研究进展作一综述。

关键词：棘阿米巴副衣原体病原学分离培养

衣原体包涵体膜蛋白的结构特征及生物学功能研究进展

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中国人兽共患病学报 2019年第4期35卷 338-344页

作者：肖健王川吴移谋南华大学病原生物学研究所衡阳421001

包涵体膜蛋白是一类由衣原体分泌并被转运至包涵体膜的衣原体特异性蛋白。进入宿主细胞后,衣原体即可分泌大量蛋白对包涵体膜进行修饰,这类蛋白位于衣原体-宿主细胞界面处,因此可在维持包涵体稳定性、衣原体营养摄取及与宿主细胞发生相... 详细信息

包涵体膜蛋白是一类由衣原体分泌并被转运至包涵体膜的衣原体特异性蛋白。进入宿主细胞后,衣原体即可分泌大量蛋白对包涵体膜进行修饰,这类蛋白位于衣原体-宿主细胞界面处,因此可在维持包涵体稳定性、衣原体营养摄取及与宿主细胞发生相互作用的过程中发挥重要功能。本文就包涵体膜蛋白的结构特征及生物学功能研究进展进行了综述,以期更深入了解这种衣原体效应蛋白。

关键词：衣原体包涵体膜蛋白生物学功能

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内盖夫西门坎菌的研究现状与展望

中国人兽共患病学报 2021年第1期37卷 65-70页

作者：卢思敏袁帅吴移谋南华大学病原生物学研究所衡阳421001

内盖夫西门坎菌隶属于衣原体目西门坎菌科,是一种能广泛寄生于节肢动物、哺乳动物细胞和阿米巴虫等各种生物的新型衣原体。与目前已知衣原体相比,其具有许多独特的生物学特点,如西门坎菌囊泡与内质网密切相关、23S rRNA上存在Ⅰ型内含... 详细信息

内盖夫西门坎菌隶属于衣原体目西门坎菌科,是一种能广泛寄生于节肢动物、哺乳动物细胞和阿米巴虫等各种生物的新型衣原体。与目前已知衣原体相比,其具有许多独特的生物学特点,如西门坎菌囊泡与内质网密切相关、23S rRNA上存在Ⅰ型内含子等。血清学和PCR结果显示,此菌在人群中广泛分布,人类可能通过接触携带病原菌的阿米巴虫、节肢动物或饮用被污染的水后被感染,从而引发婴儿细支气管炎、成人社区获得性肺炎和慢性阻塞性肺疾病急性加重等呼吸道疾病的发生和发展。因此,内盖夫西门坎菌的研究为临床检测和防治提供重要线索和途径。本文就内盖夫西门坎菌研究现状作一综述。

关键词：内盖夫西门坎菌生物学特征致病性和致病机制实验室诊断

吡格列酮干预下糖尿病大鼠肾组织TLR4和PPARγ的相关性研究

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中国免疫学杂志 2014年第3期30卷 383-387页

作者：余益本吴移谋文格波湖北医药学院附属东风医院老年病科十堰442000 南华大学病原生物学重点实验室衡阳421001 南华大学国家临床医学研究中心衡阳421001

目的：研究吡格列酮对糖尿病大鼠肾组织TLR4及PPARγ表达变化规律及其意义。方法：取57只5周龄180—200g的清洁级雄性SpragueDawley（SD）大鼠，随机分为正常对照组（A组，n=9）和造模组（n=48）。造模组给予腹腔注射链脲佐菌素（Strept... 详细信息

目的：研究吡格列酮对糖尿病大鼠肾组织TLR4及PPARγ表达变化规律及其意义。方法：取57只5周龄180—200g的清洁级雄性SpragueDawley（SD）大鼠，随机分为正常对照组（A组，n=9）和造模组（n=48）。造模组给予腹腔注射链脲佐菌素（Streptozoein，STZ，50mr：／kg）诱导大鼠成糖尿病模型（其中3只造模未成功）。将造模成功的40只大鼠随机分为：吡格列酮5mg／（kg·d）（B组，n=9），吡格列酮10mg7（kg·d）组（C组，n=9），吡格列酮5mg／（kg·d）＋TLR4特异性抑制剂Eritoran5mg／（kg·d）（D组，n=9），吡格列酮10mg／（kg·d）＋TLR4特异性抑制剂Eritoran5mg／（kg·d）（E组，n=9），未干预组（F组，n=9），于第8周末，取肾组织予以HE染色，HE染色后用光镜观察肾组织形态学改变，采用Real—timePCR检测肾组织中TLR4和PPARγ的mRNA表达水平。结果：与A组相比，B、C、D、E组糖尿病大鼠肾组织TLR4的表达明显增加，PPARγ表达明显减弱，比较存在差异（P〈0．05）；与F组相比，B、C、D、E组大鼠肾组织TLR4的表达亦减弱，PPARγ表达增加，比较存在差异（P〈0．05）；C组与B组比较，糖尿病大鼠肾组织TLR4的表达减弱，PPA脚表达稍增强，前后比较无差异（P〉0．05）；E组与D组比较，肾组织TLR4的表达均明显减弱，PPARγ表达均明显增强，但无统计学差异（P〉0．05）；Pearson相关性分析提示糖尿病大鼠肾组织TLR4与PPARγ表达呈负相关，相关系数为-0．962（P〈0．05）。结论：吡格列酮对糖尿病肾病具有保护作用，部分作用机制可能是通过抑制TLR4的表达，促进PPAR叮表达，抑制炎症反应而实现的。

关键词：吡格列酮过氧化物酶体增殖物激活受体γ Toll样受体4 糖尿病肾病炎症反应