酶联免疫吸附试验两步法在HIV抗体检测中的应用

目的探讨酶联免疫吸咐试验（ELISA）双抗原夹心一步法和二步法检测人类免疫缺陷病毒（HIV）抗体的不同效果。方法采集2005年10月-2008年10月本市中心血站无偿献血者血液标本48463例,用双抗原夹心ELISA一步法和二步法试剂检测,初筛阳性标本,采用免疫印迹法进行确证。结果检测48463份标本,一步法初筛阳性87例,确证阳性4例,假阳性率0．17％;二步法初筛阳性24例,确讧阳性4侧,假阳性率0．05％,两法的假阳性率比较有显著性差异（P〈0．01）。结论酶联免疫吸咐试验二步法检测人类免疫缺陷病毒抗体的效果优于一步法,更适合于人类免疫缺陷病毒检测的初筛试验。     

扬州地区男性STD6种支原体感染状况初步研究

目的：探讨扬州地区男性STD6种致病性支原体感染状况。方法：应用nestedPCR(nPCR)技术对56例男性SID者的尿道拭子进行解脲脲原体（Uu)，人型支原体（Mh)，肺炎支原体（Mpn)，生殖支原体（Mg)。发酵支原体（Mf)，穿通支原体（Mpe)等6种支原体联合检测。结果：Uu,Mh,Mpn,Mg,Mf,Mpe阳性率分别为69.6%,35.7%,28.6%,16.1%,3.6%,3.6%；单种支原体到4种支原体联合检测。结果：Uu,Mh,Mpn,Mg,Mf,Mpe阳性率分别为69.6%,35.7%,28.6%,16.1%,3.6%,3.6%；单种支原体到4种支原体合并感染率分别为35.7%,21.4%,14.3%和8.9%；6种支原体均为阴性占19.6%。结论：扬州地区男性STD者6种支原体感染率较高。本文并就支原体感染对妊娠等的影响进行了讨论。     

女性生殖道感染治疗前后解脲支原体荧光定量PCR检测的假阳性分析

目的探讨荧光定量PCR（FQ—PCR）检测解脲支原体的假阳性率，为客观分析其结果提供依据。方法采用FQ—PCR及培养法同时检查了70例无症状妇女及163例生殖道感染患者治疗前后宫颈分泌物中解脲支原体。结果无症人群FQ—PCR检测阳性率（42．9％），明显高于培养阳性率（11．4％）（P＜0．01）；生殖道感染患者FQ—PCR检测阳性率（66．9％），高于培养阳性率（45．4％）（P＜0．05）；治疗10天后FQ—PCR检测阳性率（40％），高于培养阳性率（10．8％）（P＜0．01）。结论FQ—PCR检测女性生殖道解脲支原体有一定的假阳性，应用时应客观分析其结果。     

生殖道支原体与女性不孕不育和抗子宫内膜抗体的关系

目的研究女性不孕不育和生殖道解脲支原体（U．urealyticum，UU）、人型支原体（M．hominis，MH）和抗子宫内膜抗体（Antiendometrium antibody，AEMAb）的关系。方法采用培养法分别对108例原发性和96例继发性不孕不育患者宫颈分泌物进行UU，MH检测；采用ELISA检测血清中AEMAb。结果原发性和继发性不孕不育组UU，MH感染率及AEMAb阳性率与对照组比较差异均有显著性（P〈0．00167=。不孕组中支原体感染阳性患者AEMAb阳性率明显高于支原体阴性患者（P〈0．05）。结论女性不孕不育与生殖道支原体感染和AEMAb的产生有密切关系，为临床诊断和合理治疗不孕不育患者提供了理论依据。     

不孕不育夫妇生殖道解脲支原体、沙眼衣原体感染及抗精子抗体、抗子宫内膜抗体关系探讨

目的探讨不孕不育夫妇生殖道解脲支原体（UU）、沙眼衣原体（CT）感染及抗精子抗体（AsAb）、抗子宫内膜抗体（EMAb）间的关系。方法采用PCR荧光定量法对120对不孕不育夫妇进行生殖道分泌物UU、CT检测,与对照组比较,同时采用酶联免疫法（ELISA）对不孕不育夫妇血清中AsAb和EMAb进行检测,分析比较感染组与未感染组AsAb和EMAb阳性率。结果120对不孕不育患夫妇中,男性：UU感染率为25．8％、CT感染率为24．2％,UU与CT混合感染率为10．0％,与对照组男性比较差异有统计学意义P〈0．05,女性：UU感染率为33．3％。CT感染率为26．7％,UU与CT混合感染率为11．7％,与对照组女性比较差异均有统计学意义P〈0．05。在不孕不育夫妇感染组中,AsAb阳性率为32．3％,与未感染组比较差异有高度显著性P〈0．01,EMAb阳性率为10．8％,与未感染组比较差异有统计学意义P〈0．05。结论生殖道UU、CT感染是造成不孕不育的原因之一,AsAb、EMAb的产生与生殖道UU、CT感染有关。     

生殖道感染与先兆流产关系的临床研究

目的 探讨生殖道念珠菌、解脲支原体（UU）和沙眼衣原体（CT）感染与先兆流产之间的关系。方法 先兆流产患者70例作为研究组，同期随机抽取60例正常早孕妇女作为对照组，已婚未育妇女50例作为正常组。对三组妇女进行了白带常规的检测，其中念珠菌检测采用直接镜检法。同时使用培养法检测UU，聚合酶链反应（PCR）法检测CT。结果 研究组、对照组、正常组妇女白带常规中白细胞超标率分别为60％、36．7％、12％，清洁度超标率分别为57．1％、33．3％、14％，念珠菌阳性率分别为22．9％、26．7％、10％，UU阳性率分别为32．9％、40％、38％；CT阳性率分别为21．4％、13．3％、3．3％；CT＋UU阳性率分别为11．4％、3．3％、4％。结论 生殖道CT和UU＋CT混合感染与先兆流产密切相关。     

广东地区2126例泌尿生殖道支原体感染及药敏结果分析

目的了解广东地区泌尿生殖道支原体感染的情况以及对药物的敏感性。方法采用支原体培养、鉴定、计数及药敏一体化试剂盒对2126例泌尿生殖系感染患者进行检测及药物敏感性试验。结果2126例中支原体阳性1080例，阳性率为50．80％。其中解脲支原体（Uu）、人型支原体（Mh）及解脲和人型支原体（Uu＋Mh）混合感染的阳性率分别为37．53％（798例）、2．72％（58例）、10．54％（224例）。Uu对12种抗菌药物敏感性依次是克拉霉素（89．15％）、美满霉素（87．00％）、强力霉素（86．00％）和四环素（81．80％），Mh敏感性较高的依次是强力霉素（95．20％）、美满霉素（95．20％）和交沙霉素（84．10％），Uu＋Mh对12种抗生素的敏感性均较差。结论广东地区支原体感染主要以Uu为主。其次是Uu＋Mh混合感染；Uu、Mh和Uu＋Mh三者对抗生素的敏感性存在一定的差异，临床上应根据支原体培养及药敏试验结果合理用药，以最大限度地控制新耐药菌株的产生。     

重庆地区PCR检测慢性泌尿生殖道炎症患者的淋病以球菌,沙眼…

为明确慢性泌尿生殖道炎症的病原体种类我们对重庆地区８７例男女患者采用ＰＣＲ技术检测。结果５４例男性患者尿道分泌物中淋球菌（ＮＧ）、沙眼衣原体（ＣＴ）及解脲支原体（ＵＵ）的检出阳性率分别为５９．２％、３３．３％和７．４％。３３例女性患者宫颈分泌物中的ＮＧ、ＣＴ和ＵＵ的检出阳性率为５４．５％、４５．５％和３９．４％。在５４例男性分泌物中同时检出ＮＧ与ＣＴ合并感染的１０例，检出ＮＧ与ＵＵ合并感染的为１例     

继发不育男性泌尿生殖道支原体感染状况与耐药性分析及其临床应用研究

目的探讨继发不育男性患者泌尿生殖系支原体的感染状况及其耐药性。方法采用支原体分离培养鉴定与药敏试剂盒，对继发不育男性患者泌尿生殖系分泌物进行支原体分离培养鉴定与药敏测试。结果572例继发不育男性患者，泌尿生殖系支原体培养阳性278例，阳性率为48．6％，其中Uu占90．3％（251／278）；Mh占1．4％（4／278）；Uu合并Mh占8．3％（23／278）。药敏试验表明：喹诺酮类耐药率最高，四环素类和大环内酯类耐药率最低。结论支原体感染是男性继发不育的重要危险因素，在支原体感染的治疗过程中，我们应根据药敏测试结果选择敏感抗生素。     

用套式PCR对泌尿生殖道生殖支原体感染现状的检测

了解泌尿生殖道生殖支原体的感染状况，方法：采用套式ＰＣＲ对临床正常者４６例和泌尿生殖道炎症患者１４２例，进行生殖支原体的检测。结果；１临床正常男女性泌尿生殖２道生殖支原体的检出率为４．３％。２．泌尿生殖道炎症患者的检出率为４０．８％，其中合并ＮＧ，ＣＴ或ＵＵ率占２３．２％，单纯生殖支原体感染率占１７．６％。     

Identification and characterization of immunogenic proteins of mycoplasma genitalium

Mycoplasma genitalium causes nonchlamydial nongonococcal urethritis. M. genitalium was detected by PCR in 17 urethral swabs obtained from 99 men with and without urethritis (J. S. Jensen, R. Orsum, B. Dohn, S. Uldum, A. M. Worm, and K. Lind, Genitourin. Med. 69:265-269, 1993), and later, four M. genitalium strains were isolated (J. S. Jensen, H. T. Hansen, and K. Lind, J. Clin. Microbiol. 34:286-291, 1996). The objective of this study was to characterize immunogenic proteins of M. genitalium by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting by using a hyperimmune rabbit serum against M. genitalium G37, determine their identity by mass spectrometry, and develop an M. genitalium-specific enzyme-linked immunosorbent assay (ELISA) free from cross-reactivity with M. pneumoniae antibodies. Using recombinant fragments of the C-terminal part of MgPa (rMgPa), we developed a specific ELISA for detection of M. genitalium antibodies. This antigen did not bind M. pneumoniae antibodies. Using serum samples from the 99 men with and without urethritis, we found that 26 had immunoglobulin G (IgG) antibodies to M. genitalium. There was a strong statistically significant correlation between PCR and IgG antibodies to M. genitalium (odds ratio [OR], 5.9; 95% confidence interval [CI], 2.3 to 21.5; P = 0.002). Furthermore, men with recurrent urethritis were more likely to have antibodies to M. genitalium than were those without recurrent urethritis (OR, 4.0; 95% CI, 1.1 to 14.5; P = 0.0383) and they had significantly higher antibody titers. By use of the rMgPa ELISA, this study further substantiates the importance of M. genitalium as a cause of male urethritis.     

The use of enzyme-linked immunosorbent assay for detection of Mycoplasma hominis antibodies in infertile women serum samples

BACKGROUND: Besides Chlamydiae trachomatis and Mycoplasma genitalium, Mycoplasma hominis may also cause infertility due to damage of the Fallopian tubes. Therefore serum samples from infertile women were analyzed for antibodies to M. hominis. METHODS: Sera from 304 infertile women were investigated for seropositivity to M. hominis by immunoblotting and a developed ELISA. Women were classified into groups based on the type of infertility: infertile due to lack of passage in Fallopian tubes (TFI, tubal factor infertility), an infertile male partner (MFI, male factor infertility) and unexplained infertility (UFI, unexplained factor infertility). Three M. hominis isolates were used in the immunoblotting analysis and clear differences in patient immunoprofiles were observed between two isolates. For the ELISA we used a mixture of Triton X-114 extracted membrane proteins from those two M. hominis isolates as antigen. RESULTS: Ninety-seven sera (32%) were seropositive to M. hominis when tested by the ELISA. There was a significant correlation between TFI and seropositivity to M. hominis (P = 0.0015, OR = 2.21, CI = 1.35-3.61). We compared the seropositivity of 304 patients to M. hominis with the presence of antibodies against two other bacteria Chlamydiae trachomatis and Mycoplasma genitalium and there was no statistical correlation between those bacteria and M. hominis. CONCLUSION: Our results indicate that M. hominis may be an independent predictor of TFI.     

Comparison of enzyme-linked immunosorbent assay (ELISA) and complement fixation test for detection of Mycoplasma pneumoniae antibodies.

An enzyme-linked immunosorbent assay (ELISA) for the detection of IgG and IgM antibodies against Mycoplasma pneumoniae, performed with commercial antigen and reagents, is compared with the complement fixation test (CF) in a serological study of 209 human sera. Concordant results were usually obtained by CF test and by IgG ELISA in sera from patients with recent M pneumoniae infection. In contrast, when used for an immunological survey of a general population, approximately 27% of the sera negative in the CF test were positive for IgG by the ELISA, and sera with low CF titres were found to have a broad range of IgG titre by the ELISA. This may be due to the greater sensitivity of the ELISA technique and/or to different types of antibody measured by both tests. IgM was detected by ELISA in sera from all patients with recent M pneumoniae infection diagnosed on the basis of clinical findings and by CF assay. Occasionally false-positive IgM antibodies were due to rheumatoid factor (RF); this potential interference necessitates routine testing of IgM antibody positive sera for RF.     

Serological investigation of Mycoplasma genitalium in infertile women

BACKGROUND: The role of Mycoplasma genitalium in the pathogenesis of pelvic inflammatory disease has not been characterized. METHODS: Sera from 308 infertile women were investigated for antibodies to M. genitalium by immunoblotting. Women with tubal factor infertility (TFI) made up 132 of the patients, 67 of the women had an infertile male partner and 109 were infertile for unknown reasons. RESULTS: Of the TFI patients 29 (22.0%) were seropositive to the major adhesin, MgPa, of M. genitalium versus 11 (6.3%) in the group of women with normal tubes. No cross-reactions between MgPa and P1 of the related Mycoplasma pneumoniae were found. Besides, MgPa positive sera were confirmed by immunoblotting using a cloned fragment of the C-terminal part of MgPa specific to M. genitalium. Chlamydia trachomatis is known to be able to cause infertility as a result of salpingitis. Therefore, the sera were tested against C. trachomatis using a commercial ELISA test. Seventy-five (56.8%) of the TFI patients were seropositive to C. trachomatis. Eight (27.6%) TFI patients seropositive to MgPa were negative to C. trachomatis. CONCLUSIONS: This study indicates that M. genitalium may be an independent risk factor in the development of an inflammatory process leading to scarring of the uterine tubes in women and thereby causing infertility.     

Immunological cross-reactivity of a Mycoplasma pneumoniae membrane-associated protein antigen with Mycoplasma genitalium and Acholeplasma laidlawii.

A murine monoclonal antibody, OC2F5, reacts with a Mycoplasma pneumoniae antigen with an approximate Mr of 43,000. This antigen is trypsin and proteinase K sensitive and partitions in the detergent phase of a Triton X-114 solution. The monoclonal antibody cross-reacts with an antigen from both Mycoplasma genitalium and Acholeplasma laidlawii with a similar molecular weight. This cross-reactivity should be considered in the development of M. pneumoniae antigen detection systems based on the use of antibodies directed to this protein antigen.     

Diagnostic use of polyclonal antibodies raised in mouse ascitic fluid in bancroftian filariasis

Polyclonal antibodies were produced against Brugia malayi adult antigens (BmA (PBS) SAg and BmA (SDS) SAg) in mouse ascitic fluid by immunising Balb/c mice intraperitoneally with high ratio of adjuvant to immunogen. The diagnostic use of these antibodies in detecting circulating filarial antigen in bancroftian filariasis was studied by sandwich enzyme-linked immunosorbent assay (sandwich ELISA) using stick assay system. Both antibodies raised against PBS and SDS soluble antigens were found to be equally sensitive and relatively specific in detection of circulating filarial antigen. When anti BmA (PBS) SAg antibody was used in sandwich ELISA, 90% of microfilaraemic sera, 30-40% of acute and sub acute filarial sera, 20% of chronic filarial sera, 7% of endemic normal sera and none of 15 non-endemic normal sera were positive for filarial antigen. Using anti BmA (SDS) Sag antibody, 93% of microfilarial sera, 40% of acute and sub acute filarial sera, 20% of chronic filarial sera and none of 15 endemic and non-endemic normal sera showed the presence of filarial antigen. The filarial antigen detection using anti BmA S Ag antibodies produced in mouse ascitic fluid in sandwich ELISA may be useful in detection of active stage (microfilaraemia) of infection.     

抗日本血吸虫SEA特异性IgY应用于循环抗原检测的研究

本研究应用抗日本血吸虫可溶性虫卵抗原(SEA)的鸡卵黄免疫球蛋白(IgY)建立敏感、特异的检测循环抗原的双抗体夹心酶联免疫吸附试验( ELISA).用SEA皮下多点注射法免疫海蓝鸡,水稀释法制备IgY抗体,以辣根过氧化物酶标记纯化的IgY抗体(IgY-E)和兔抗IgG抗体(IgG-E)分别作为检测抗体,IgY抗体和兔抗...     

应用双抗原夹心ELISA法筛查献血员梅毒螺旋体抗体

目的：优选一种适宜于献血员梅毒筛查的试验方法。方法：采用检测梅毒特异性抗体的双抗原夹心ELISA法对献血员进行抗体检测，并与RPR检测结果进行比较，对两种方法检测结果不一致的标本，再用TPHA法进行确证。结果：ELISA法阳性检出率0．36％(41／1271)、RPR法阳性检出率0．26％(29／11271)。ELISA法与TPHA法总符合率97．5％(40／41)、RPR法与TPHA总符合率63．41％(26／41)。结论：ELISA法优于RPR法，具有较高的灵敏度和特异性，适宜于献血员的筛查．有利于控制和减少梅毒的输血传播。     

Comparison of host responses after intranasal infection of guinea-pigs with Mycoplasma genitalium or with Mycoplasma pneumoniae

Guinea-pigs were infected intranasally with Mycoplasma genitalium or Mycoplasma pneumoniae. The lung lesions produced by the two mycoplasmas were comparable in extent and histological pattern. Sera of both animal groups taken 2 weeks after infection reacted strongly in the complement fixation test with the M. pneumoniae glycolipid extract. In an ELISA using the respective adherence proteins (P1-protein of M. pneumoniae and MgPa of M. genitalium), strong specific activity, but also considerable cross-reactions were found. Epitope analysis by using overlapping octapeptides of a P1-region immunologically active in human M. pneumoniae infections and of the corresponding MgPa-region revealed six common epitopes but also one M. genitalium and two M. pneumoniae specific determinants. For analysis of a possible pathogenicity of M. genitalium in the human respiratory tract species-specific tests have to be developed.     

Application of an indirect immunofluorescence test for detection of Mycoplasma pneumoniae in respiratory exudates.

We prepared polyclonal antibody specific to Mycoplasma pneumoniae and examined the conditions influencing the ability of an indirect immunofluorescence test to detect the specific antigen in respiratory exudates. The antibody did not cross-react with normal human serum or with respiratory exudates from 10 healthy persons. Cross-reactivity of the antibody with species of mycoplasmas other than M. genitalium was fully diminished when absorbed with horse serum and yeast extract, components of the culture medium. Though the absorbed antibody cross-reacted with M. genitalium, the titer was significantly lower than when tested against M. pneumoniae. Two types of antigen-specific fluorescence were observed in clinical specimens: one is large or small fluorescent granular aggregates found in mucus, and the other is fine fluorescent particles diffused on the entire surface of small epithelial cells. Throat smears from 49 patients with serologically confirmed M. pneumoniae infections were examined by our indirect immunofluorescence method. Positive results were obtained in 42 cases, many of which were positive before a rise in serum antibody titer could be demonstrated, indicating that the method is useful for a preliminary diagnosis at an early stage of the infection.