聚合酶链反应检测重型肝炎患者血清HBVDNA

采用ＰＣＲ法检测４８例重型肝炎患者血清中ＢＨＶＤＮＡ存在情况，并与斑点杂交检测进行比较，结果表明：ＰＣＲ检测３９例阳性，检出率为８１．３％（３９／４８）．ＨＢｓＡｇ（＋）、ＨＢｅＡｇ（＋）组ＰＣＲ检测均阳性（８／８），ＨＢｓＡｇ（＋）、ＨＢｅＡｇ（－）组阳性率为８８．０％（２２／２５），抗体阳性组检出率为７２．７％（８／１１），检测灵敏度为１．０ｆｇ；斑点杂交检测１５例阳性，阳性率３０．２％（１５／４８），各组斑点杂交阳性率均低于ＰＣＲ法．提示：（１）大多数重型肝炎患者血清中存在低滴度的病毒颗粒，对重肝发病有一定作用；（２）大部分ＨＢｅＡｇ阴性，抗体阳性的重型肝炎患者血清仍有传染性，应考虑抗病毒治疗．     

结,直肠癌上皮生长因子受体与雌激素受体的表达及相互关系

用免疫组化ＡＢＣ法检测２７例结、直肠癌的上皮生长因子受体（ＥＧＦＲ）和雌激素受体（ＥＲ）。结果：ＥＧＦＲ阳性１６例（５９．３％），ＥＲ阳性８例（２９．６％），ＥＧＦＲ、ＥＲ均阳性７２例（２５．９％），均显著性高于正常对照组（Ｐ〈０．０５），ＥＧＦＲ、ＥＲ表达与Ｄｕｄｅｓ分期有关，ＥＧＦＲ的阳性率在癌组织中显著高于ＥＲ（Ｐ〈０．０５）。１１例淋巴结转移腺癌中，ＥＧＦＲ阳性８例（７２．７％），ＥＲ阳性     

非霍奇金淋巴瘤中EB病毒BNLF1片段及其表达产物的检测及意义

为研究ＥＢ病毒，与非霍奇金淋巴瘤（ＮＨＬ）的关系，采用ＰＣＲ，原位杂交，ＬＳＡＢ（ｌａｂｅｌｌｉｎｇｓｔｒｅｐｔａｖｉｄｉｖｉｎｂｉｏｔｉｎ）免疫组化技术，检测３２例ＮＨＬ和３３例反应性增生淋巴结中的ＥＢＶ－ＢＮＬＦ１的基因片段及其表达产物潜伏膜蛋白（ＬＭＰ１），ＮＨＬ中ＥＢＶ－ＢＮＬＦ１ＰＣＲ扩增阳性率为５３％，明业高于反应性增生淋巴结组２７％（Ｐ〈０．０５），其中５例Ｔ细胞淋巴瘤全部阳性，１７     

PCR技术在霍乱监测中的应用研究

为探讨ＰＣＲ检测技术在霍乱监测中的应用价值。我们于１９９６年７月至１９９８年１１月霍乱监测工作中,应用ＰＣＲ技术与培养法平行检测各类监测标本６７８份,结果ＰＣＲ检出阳性率９．４４％显著高于培养法的５．７５％（ｘ＾２＝６．５６,ｐ＝０．０１０４〈０．０５）。其中健康接触者ＰＣＲＳ同率为１３．７３％,显著高于培养法的１．９６％（ｘ＾２＝４．８８,ｐ〈０．０５４）;水标本ＣＲ检出率１０．８％也显著高于培     

非霍奇金淋巴瘤中EB病毒BNLF_1片段及其表达产物的检测及意义

为研究ＥＢ病毒与非霍奇金淋巴瘤（ＮＨＬ）的关系，采用ＰＣＲ、原位杂交、ＬＳＡＢ（ｌａｂｅｌｉｎｇｓｔｒｅｐｔａｖｉｄｉｖｉｎｂｉｏｔｉｎ）免疫组化技术，检测３２例ＮＨＬ和３３例反应性增生淋巴结中的ＥＢＶＢＮＬＦ１基因片段及其表达产物潜伏膜蛋白（ＬＭＰ１）。ＮＨＬ中ＥＢＶＢＮＬＦ１ＰＣＲ扩增阳性率为５３％，明显高于反应性增生淋巴结组２７％（Ｐ＜０．０５），其中５例Ｔ细胞淋巴瘤全部阳性。１７例ＰＣＲ阳性的ＮＨＬ中５例原位杂交阳性。ＬＭＰ１检测仅２例阳性。结果表明本组病例半数以上ＮＨＬ中有ＥＢＶ感染，可能与肿瘤的发生发展有一定关系。     

PCR法检测泌尿道感染患者沙眼衣原体与解脲脲原体的研究

用聚合酶链反应（ＰＣＲ）法，对门诊的５２８例初诊为泌尿道感染患者进行了沙眼衣原体（Ｃｔ）与解脲脲原体（Ｕｕ）检测。结果显示：１７９例为淋球菌阳性，其中ＰＣＲ检测Ｃｔ阳性７１例，阳性率为３９．７％，Ｕｕ阳性７７例，阳性率为４３．０％；在３９４例非淋菌性尿道炎（ＮＧＵ）中，Ｕｕ阳性率为５３．３％（１８６／３４９），Ｃｔ阳性率为２７．２％（９５／３４９）。本研究表明应用ＰＣＲ检测Ｃｔ与Ｕｕ具有敏感性高、     

结、直肠癌上皮生长因子受体与雌激素受体的表达及相互关系

用免疫组化ＡＢＣ法检测Ｚ７例结、直肠癌的上皮生长因子受体（ＥＧＦＲ）和雌激素受体（ＥＲ）。结果：ＥＧＦＲ阳性１６例（５９．３％），ＥＲ阳性８例（２９．６％），ＥＧＦＲ、ＥＲ均阳性７例（２５．９％），均显著性高于正常对照组（Ｐ＜０．０５），ＥＧＦＲ、ＥＲ表达与Ｄｕｄｅｓ分期有关，ＥＧＦＲ的阳性率在癌组织中显著高于ＥＲ（Ｐ＜０．０５）。１１例淋巴结转移腺癌中，ＥＧＦＲ阳性８例（７２．７％），ＥＲ阳性７例（６３．６％），两者均阳性５例（４５．５％）。结果提示：ＥＧＦＲ、ＥＲ与结、直肠癌有关，表皮生长因子在肿瘤的发生发展中的作用远较雌激素重要。     

急性淋巴细胞白血病微量残留病的PCR监测

①目的探讨急性淋巴细胞白血病（ＡＬＬ）微量残留病（ＭＲＤ）检测的临床意义。②方法应用筑巢式多聚酶链反应（ｎｅｓｔｅｄＰＣＲ）对３２例ＡＬＬ进行Ｔ细胞受体（ＴＣＲ）Ｖδ２Ｄδ３基因重排检测。③结果１８例Ｂ系ＡＬＬ中有１５例（７２．２％）、４例Ｔ系ＡＬＬ中有１例（２５．０％）存在ＴＣＲＶδ２Ｄδ３基因重排。同时对１６例进行３９例次微量残留病动态监测，其中６例ＭＲＤ－ＰＣＲ阴性病人随访５．１７～１６．１７年，无１例复发；１０例ＭＲＤ－ＰＣＲ阳性者，２例分别于阳性后０．２５，１．００年骨髓复发，１例有复发倾向，余７例随访４．３３～６．２５年无复发。④结论ＭＲＤ－ＰＣＲ阴性者预后良好，可望长期生存，治疗时应以ＭＲＤ－ＰＣＲ转阴为停止化疗的可靠指标，对ＭＲＤ－ＰＣＲ阳性者应定期监测ＭＲＤ变化，结合病人情况进行综合评价。     

阴道加德纳菌致病株的检测

应用聚合酶链反应（ＰＣＲ）法检测阴道加德纳菌（Ｇ．Ｖａｇ）致病株,探讨其临床诊断价值。在ＩＴＳ－２３ｓｒＲＮＡ基因上设计一对特异性引物Ｐ１／Ｐ２,建立ＰＣＲ反应体系,特异地扩增Ｇ．Ｖａｇ致病株。结果Ｇ．Ｖａｇ扩增片段４３３ｂｐ,１４１例有临床症状者检出３６例阳性,阳性率２５５％,１２９例无症状对照组检出６例阳性,阳性率４６％,两者比较差异有显著性。该ＰＣＲ反应体系特异地检测Ｇ．Ｖａｇ致病株,为临床诊断及其致病性的研究提供了新的检测方法     

PCR法检测泌尿道感染患者沙眼衣原体与解脲脲原体的研究

用聚合酶链反应（ＰＣＲ）法，对门诊的５２８例初诊为泌尿道感染患者进行了沙眼衣原体（Ｃｔ）与解脲脲原体（Ｕｕ）检测。结果显示：１７９例为淋球菌阳性，其中ＰＣＲ检测Ｃｔ阳性７１例，阳性率为３９．７％，Ｕｕ阳性７７例，阳性率为４３．０％；在３９４例非淋菌性尿道炎（ＮＧＵ）中，Ｕｕ阳性率为５３．３％（１８６／３４９），Ｃｔ阳性率为２７．２％（９５／３４９）。本研究表明应用ＰＣＲ检测Ｃｔ与Ｕｕ具有敏感性高、特异性强等优点，能为临床泌尿道感染病因学诊断与治疗提供定性依据     

Clinical,radiological and molecular diagnosis correlation in serum samples from patients with osteoarticular tuberculosis

ObjectiveTo assess the role of polymerase chain reaction (PCR) in serum samples, in the diagnosis of osteoarticular tuberculosis (OTB) in a setting where only clinical and imaging diagnoses determine the treatment.MethodsA total of 44 consecutive serum specimens were collected from clinically suspected OTB patients, based on clinical and radiological [X-ray or magnetic resonance imaging/computed tomography] features. They were screened by in-house nested PCR. In addition, a few specimens were examined by Gram stain, acid-fast bacilli stain, histopathology and routine bacterial culture. A total of 39 specimens were collected from patients suffering from other bone diseases of nontuberculous origin and included as negative controls.ResultsOf the 44 clinically suspected OTB patients, in-house nested PCR was positive in 40 (91%) cases; PCR was negative in 38 (97%) negative controls. Sensitivity and specificity of our in-house nested PCR was 90.9% and 97.4%, respectively. The PCR report was available within 48 h. It was possible to standardize serum PCR technique and in positive cases, a good correlation was observed in terms of an adequate treatment response.ConclusionsNested PCR in serum samples is a rapid, highly sensitive and specific modality for OTB detection. PCR should be performed in addition to clinical evaluation, imaging studies, acid-fast bacilli staining, culture and histopathology diagnosis, if possible.     

应用聚合酶链反应快速诊断巨细胞病毒感染

用聚合酶链反应（ＰＣＲ）技术检测学龄前儿童尿标本中的巨细胞病毒（ＨＣＭＶ），并和常规细胞培养（ＣＣＣ）、早期抗原检测（ＤＥＡ）两法进行了比较。结果表明，１０５份尿标本中有４１份ＣＣＣ出现ＨＣＭＶ特征性细胞病变（ＣＰＥ），其中３８份标本ＰＣＲ检测为阳性，敏感性为９２．７％；其余６４份ＣＣＣ阴性，有６份ＰＣＲ检测阳性，特异性为９０．６％。与ＤＥＡ相比，ＰＣＲ检测的特异性和敏感性分别为９３．６％，９５．２％；提示ＰＣＲ检测临床标本中ＨＣＭＶ，不仅敏感特异，而且与ＣＣＣ，ＤＥＡ两法相比快速简便。     

Nested polymerase chain reaction for early diagnosis of typhoid fever

Typhoid fever, caused by Salmonella typhi, is an important cause of morbidity and mortality in many developing countries. A rapid and sensitive method for the detection of S. typhi is essential for early diagnosis. This was a study to prospectively evaluate the sensitivity and specificity of nested polymerase chain reaction (PCR) to identify the S. typhi using flagellin gene related primers. The study was carried out in the department of Microbiology, Mymensingh Medical College, Mymensingh between July, 2010 and June, 2011, including 82 individuals of different age and sex. Of them, 62 were clinically suspected cases of typhoid fever and remaining 20 were apparently healthy controls. Cultures as well as PCR of blood specimens were performed for each of the cases. Among the 62 suspected typhoid fever cases, 8(12.9%) were blood culture positive and 55(88.7%) were PCR positive for S. typhi. All culture positive cases were positive by PCR and among 54 culture negative cases, 47(87%) were positive by PCR. Neither of the healthy controls was positive by PCR or blood culture. The sensitivity, specificity, positive predictive value and negative predictive value of PCR using blood culture as gold standard were 88.7%, 100%, 100% and 74% respectively for typhoid fever. In this study, the PCR appears highly specific, very sensitive and superior to blood culture for the early diagnosis of typhoid fever.     

免疫酶染色诊断弓形虫感染的研究

用免疫酶染色法(IEST)探讨了弓形虫感染的诊断价值。结果表明,人工感染弓形虫11份家兔和间接荧光抗体(IFAT)与间接血凝(IHA)抗体均阳性的34份人血清,阳性分别为11份和32份,4份阴性兔血清和108份抗体阴性人血清阳性分别为0份和2份,其敏感性为94.10%～100.00%,特异性为98.15%～100.00%.35份其它寄生虫感染者均未有交叉反应。11份阳性血清间隔半个月进行了4次重复试验,其阳性率及几何均数的倒数(GRMT)均无显著性差异。用IEST对健康体检者、不良生育史孕妇和智力低下儿童进行了测定,阳性率分别为8.46%,16.33%和28.26%,结果与IFAT无显著性差异。该法操作简单,值得在基层推广应用。     

淋病奈瑟菌PCR检测的临床应用研究

为评价淋病奈瑟菌ＰＣＲ检测的临床效果,建立了对标准淋病奈瑟菌的ＤＮＡＰＣＲ检测技术。结果显示,ＮＧ ＰＣＲ技术的灵敏度约为３个菌的基因组拷贝。对８种菌株进行ＮＧ ＰＣＲ,只有标准淋病奈瑟菌显示阳性,其余７株非淋球菌菌株均阴性,说明有很好的特异性。对４９例淋病患者同时进行涂片镜检法、细菌培养法和ＰＣＲ法检测的比较,其阳性检出率分别为７６．６％、５２．２％和９５．９％。提示ＮＧ ＰＣＲ方法比涂片和培养法更为敏感,且特异性强,在淋病的诊断,特别是早期诊断中有重要参考价值。     

用PCR检测消化道巨细胞病毒感染

应用多聚酶链反应（ＰＣＲ）检测５１例消化道疾病患者活检或手术标本的人类巨细胞病毒（ＨＣＭＶ）－ＤＮＡ，阳性率９．８０％；采用间接免疫荧光法随机检测其中４１例患者血清ＨＣＭＶ－ＩｇＭ。结果显示：消化道组织巨细胞病毒感染数低于血清ＨＣＭＶ－ＩｇＭ阳性数（Ｐ〈０．０１）；血清ＨＣＭＶ－ＩｇＭ检测中，恶性肿瘤和结肠炎患者ＨＣＭＶ感染均较对照组为高（Ｐ〈０．０１和Ｐ〈０．０５）。提示ＰＣＲ是消化道巨细胞病毒     

郑州市2000年夏秋季腹泻病原菌谱调查

目的：了解郑州市夏秋季感染性腹泻病原菌分布情况。方法：对2000年6至10月郑州市390例腹泻患者粪便取样进行主要病原菌的分离与鉴定，并且建立了一种快速标本处理和多重聚合酶链反应（PCR）诊断方法。结果：共分离出66株病原菌，总检出率为16．9％（66／390）。其中致病性大肠杆菌46株（11．8％），侵袭性大肠杆菌11株92．8％），志贺菌7株（1．8％），沙门菌2株（0．5％）。随机抽取67份腹泻标本，应用多重PCR检测，检出病原菌9例，高于培养法7例（P＝0．000）。结论：致病性大肠杆菌引起的婴幼儿腹泻应得到足够重视。多重PCR法对腹泻病原菌的检测具有简便、快速、特异、经济和不需培养等特点，宜于临床应用。     

PCR法检测淋球菌,沙眼衣原体及解脲支原体感染

目的：评价聚合酶链反应（ＰＣＲ）技术对性病性尿道炎及宫颈炎诊断的临床实用价值。方法：采用ＰＣＲ法、淋球菌（Ｎｇ）涂片／培养、沙眼衣原体直接免疫荧光检查（ＣｔＤＩＦＡ）和解脲支原体（Ｕｕ）培养平行检测了１１９例患者的１８９份尿道／宫颈拭子标本。结果：以Ｎｇ涂片／培养、ＣｔＤＩＦＡ和Ｕｕ培养为对比标准，ＮｇＰＣＲ、ＣｔＰＣＲ和ＵｕＰＣＲ的敏感性分别为１００％（２８／２８）、９６９％（３１／３２）和９３３％（４２／４５），特异性分别为９４５％（８６／９１）、９１９％（８０／８７）和９４６％（７０／７４）。治疗结束后１周～２周内复查仍有２５９％（２１／７０）患者ＰＣＲ未阴转。结论：ＰＣＲ技术可以敏感、特异地检测泌尿生殖道标本中的ＮｇＤＮＡ、ＣｔＤＮＡ和ＵｕＤＮＡ，具有简便、快速的优点，然而对治疗后短期内复诊患者，似不宜仅以ＰＣＲ阳性结果作为重复治疗的依据。     

PCR和培养法检测泌尿生殖道解脲支原体比较

目的 对聚合酶链反应（ＰＣＲ）和培养法检测泌尿生殖道解脲支原体（Ｕｕ）进行比较。方法 用上述二种方法同时检测１１６例患者的泌尿生殖道拭子。结果 ＰＣＲ检测阳性４６例,培养阳性４５例,培养阳性的４５例中ＰＣＲ阳性４２例。若以培养法为对照,ＰＣＲ敏感性为９３．３％,特异性为９４．４％。ＰＣＲ和增减法检测Ｕｕ的阳性率无显著差异（Ｐ〉０．０５）。结论 ＰＣＲ检测泌尿生殖道解脲支原体,敏感性高,特异性强,与     

Establishment and analysis of specific DNA patterns in 16S-23S rRNA gene spac er regions for differentiating different bacteria

Objective To establish the specific 16S-23S rRNA gene spacer regions in different bacteri a using polymerase chain reaction (PCR), restriction fragment length polymorphis m (RFLP), DNA cloning and sequences analysis. Methods A pair of primers were selected from highly conserved sequences adjacent to the 16S-23S rRNA spacer region. Bacterial DNA from sixty-one strains of standard bacteria and corresponding clinical isolates representative of 20 genera and 26 species was amplified by PCR, and further analyzed by RFLP, DNA cloning and se quences analysis. Furthermore, all specimens were examined by bacterial cultur ing and PCR-RFLP analysis. The evaluation of these assays in practical clinic practice was also discussed. Results Restriction enzyme analysis revealed one, two or three bands or more observed am ong the 26 different standard strains. The sensitivity of PCR reached 2.5 colony-forming unit (CFU), and there was no cross reaction with human genomic DNA, fungus or virus. Fourt ee n species could be distinguished immediately by PCR, while another 10 species we re further identified by Hinf Ⅰ or Alu Ⅰ digestion. The only difference betwe en K.pneumoniae and E.durans was located at the site of the 779th nucleot ide according to the sequence analysis and only XmaⅢ digestion could distingui sh one from another. Of 42 specimens from septicemic neonates, 15 were identifi e d as positive by blood culture at a rate of 35.7%. However, 27 specimens ident ified as positive by PCR, with a rate of 64.2%, a method significantly more eff ective than blood culture (P＜0.01). Of 6 cerebrospinal fluid (CSF) specim ens, one tested positi ve for S.epidermidis was also positive by PCR, two culture negative were po sitive by PCR and diagnosed as S.epidermidis according to the DNA pattern. One positive for C.neoformans was negative by PCR. The other two specimen s were negative by both PCR and culture. Conclusions The method of detecting bacterial 16S-23S rRNA spacer regions using PCR-RFLP t echniques was specific, sensitive, rapid and accurate in providing a new techniq ue for detecting pathogens in clinical bacterial infections.