结核病房空气中结核分支杆菌检测方法研究

目的探索空气中结核分支杆菌检测的有效方法。方法选用ＬＷＣ－Ⅰ型空气微生物采样器，对菌阳肺结核病房进行空气采样，３０份采样标本分别用不同方法检测结核分支杆菌。结果聚合酶链反应（ＰＣＲ）阳性１４份，阳性率４７％；Ｓｏｕｔｈｅｒｎ转印杂交阳性１８份，阳性率６０％；动物接种实验组３０只豚鼠有２只结核分支杆菌培养阳性，另１只病理组织学检查阳性，而对照组１０只豚鼠脏器培养及病理组织学检查全部阴性；罗氏培养和ＢＡＣＴＥＣ快速培养均呈阴性。结论用空气采样的方法进行空气中结核分支杆菌的监测是可行的。ＰＣＲ和Ｓｏｕｔｈｅｒｎ转印杂交最敏感，动物实验也可提供有意义的参考。     

聚合酶链反应－冷单链构象多态性快速检测结核分支杆菌rpoB基因突变

目的评估ｒｐｏＢ基因突变检测在结核分支杆菌利福平耐药性测定中的应用价值。方法采用聚合酶链反应－冷单链构象多态性（ＰＣＲ－冷ＳＳＣＰ）方法对８７株结核分支杆菌临床分离株及有药敏结果的２２份相应肺结核患者痰标本进行分析。结果ＰＣＲ扩增ｒｐｏＢ基因的敏感性为１００ｐｇＤＮＡ及５０００个菌体，属于分支杆菌特异。所有分离菌的ｒｐｏＢ基因ＰＣＲ扩增均为阳性。采用套式ＰＣＲ可使扩增敏感性提高１００倍。与传统药敏试验方法相比，ＰＣＲ－冷ＳＳＣＰ对８７株结核分支杆菌临床分离菌利福平耐药性的检测敏感性和特异性分别为８９．６％及１００％。２２份涂阳培阳痰标本中套式ＰＣＲ扩增阳性的６份均为高度利福平耐药，其ＳＳＣＰ结果也与相应分离株的药物敏感性试验相符。结论ＰＣＲ－冷ＳＳＣＰ检测结核分支杆菌ｒｐｏＢ基因突变快速、简便、易行，适用于结核分支杆菌分离株利福平耐药性的快速测定。如进一步提高引物增特异性及敏感性，该技术可望用于临床标本直接检测     

聚合酶链反应检测肺结核患者外周血单个核细胞中结核分支杆菌DNA的临床价值

应用聚合酶链反应（ＰＣＲ）技术配对检测９２例肺结核患者外周血单个核细胞内和痰标本中结核分支杆菌ＤＮＡ，同时检测３０例非结核患者外周血。结果显示：外周血和痰标本ＰＣＲ阳性率分别为７１７％和５５４％，前者明显高于后者（Ｐ＜００５）；各型肺结核外周血和痰标本ＰＣＲ阳性率不尽相同；３０例非结核患者外周血ＰＣＲ阳性３例（１０％）。认为ＰＣＲ检测肺结核患者外周血单个核细胞内结核分支杆菌ＤＮＡ是一种快速、敏感的方法，可用于肺结核早期诊断与鉴别诊断     

痰结核分支杆菌DNA扩增对肺结核疗效监测

为评价聚合酶链反应（ＰＣＲ）痰结核分支杆菌ＤＮＡ（ＴＢＤＮＡ）扩增对肺结核疗效监测的实用价值，对３１例初治涂阳肺结核患者进行了２年的随访研究。对象随机选择初治涂阳肺结核３４例，男２４例，女１０例，年龄１９～５８岁。痰涂片抗酸染色和ＰＣＲ检查阳性，胸...     

不固定化疗期短程化疗研究初探

目的研究从现行６个月短化方案进一步缩短化疗期的疗效。方法改进初治涂阳患者６个月固定化疗期，采用涂片检查并以痰菌转阴后连续３个月阴性为治愈停药标准。结果２９０例涂阳肺结核经２ＳＨＲＺ／ｘＨＲ治疗，６个月时痰菌阴转率９８．３％，２８３例阴转治愈，平均化疗期４．７个月，２年随访率９４．３％，细菌学复发率１．９％。结论采用痰涂片检查，并以痰菌转阴后连续３个月阴性为阴转治愈标准，可将化疗期平均缩至４．７个月，且治愈率高，２年细菌学复发率低，为进一步缩短化疗期进行了有意义的探索。     

核酸体外扩增技术诊断骨结核的临床研究

目的探讨核酸体外扩增（ＰＣＲ）技术在骨结核诊断中的价值。方法对６０例骨结核标本与２０例非骨结核标本分别应用ＰＣＲ、抗酸染色镜检及分离培养法进行结核分支杆菌检测。同时，分析了影响ＰＣＲ结果的有关因素与相应处理措施。结果６０例骨结核标本中三种方法的阳性检出率分别为：ＰＣＲ法８３％，镜检法３％，培养法７％。经统计学处理，Ｐ＜０．００５，ＰＣＲ法与镜检及培养法对结核分支杆菌的阳性检出率比较具有显著性差异，ＰＣＲ法明显优于镜检及培养法。２０例非骨结核标本镜检及培养法均阴性，ＰＣＲ法阳性率１０％。盲法结核分支杆菌和对照菌ＰＣＲ检测结果表明，ＰＣＲ的特异性为１００％。ＰＣＲ扩增整个过程自动化控制，可在数小时内完成。结论ＰＣＲ技术是一种快速、敏感、特异与简便的骨结核标本结核分支杆菌检测方法，对骨结核的诊断与鉴别诊断具有重要价值。     

初治痰菌阳性肺结核疗程结束时空洞或结核球内细菌学分析

目的对初治痰菌阳性肺结核病例短程化疗结束后痰菌阴转，但肺内仍存在结核球或空洞病灶的病例，经皮肺穿刺取材进行细菌学检查，评价短程化疗的疗效。方法2002年1月至2004年5月，选择初治痰菌阳性肺结核83例，患者均经过常规抗结核药物（异烟肼、利福平、乙胺丁醇、吡嗪酰胺）短程化疗，疗程末痰涂片及痰培养均为阴性，但肺部X线表现仍存在空洞或结核球，在疗程结束后1个月内，在CT引导下经皮肺穿刺对空洞壁或空洞内容物或结核球取材，进行细菌学检查，并对检查结果进行分析。结果对活检组织均进行涂片及培养检查，痰菌阴转病例中仅75例（90．4％）获得的组织标本阴性结果，8例（9.6％）获得阳性结果（其中3例为涂片阳性、培养阳性；5例为涂片阴性、培养阳性）。药敏试验结果1例对异烟肼、利福平耐药，其余7例为敏感菌株。24个月的随访结果有6例痰菌阳转。结论初治痰菌阳性空洞、结核球肺结核病例的短程化疗后，组织标本细菌学检查阳性结果的8例病例中在2年内有6例出现痰细菌学复发，提示目前普遍采用的肺结核疗效判断标准（痰菌阴转）值得商榷。     

矽肺结核及矽肺患者结核分支杆菌L型致病情况研究

目的 探讨研究矽肺结核及矽肺患者结核分支杆菌L型致病情况的价值。方法 对矽肺结核组和矽肺组患者,分别进行痰结核分支杆菌和痰结发支杆菌L型培养,并对结果进行对照观察。结果 矽肺结核组60例痰结核分支杆菌培养阳性6例,阳性率10％（同时出现结核分支杆菌L型培养阳性5例,占83％）;痰结核分支杆菌L型培养阳性28例（其中I期矽肺结核L型菌检出率为33％、Ⅱ期为7％、Ⅲ期100％）,阳性率47％（P＜0．01）矽肺组30例无一例结核分支杆菌阳性,结核分支杆菌L型阳性3例,占10％。结论 结核分支杆菌L型培养方便快速,能提高结核分支杆菌的检出率,对矽肺结核的早期诊断及减少矽肺结核复发的漏、误诊有肯定的价值、随着矽肺期别的上升,结核分支杆菌L型的检出率明显增高。     

五个月短程化疗及六个月全程间歇方案治疗菌阳肺结核的临床对照研究

目的考核利福喷丁（Ｌ）的疗效；缩短疗程或全程间歇以减少用药次数；观察全程应用吡嗪酰胺（Ｚ）对疗效及毒副反应的影响。方法以利福平（Ｒ）为对照，采用５个月疗程方案（Ⅰ组２ＳＨＲＺ／３Ｒ２Ｈ２Ｚ２，Ⅱ组２ＳＨＲＺ／３Ｌ１Ｈ２Ｚ２）、６个月全间歇方案（Ⅲ组２Ｓ３Ｈ３Ｒ３Ｚ３／４Ｌ１Ｈ２Ｚ２，Ⅳ组２Ｓ３Ｈ３Ｒ３Ｚ３／４Ｌ１Ｈ２Ｅ２），观察Ｚ的全程应用结果，巩固期以乙胺丁醇（Ｅ）为对照。３６６例初治菌阳肺结核随机分入以上４组。结果（１）３３９例完成疗程者中３２９例治疗成功，满疗程时痰菌阴转率Ⅰ～Ⅳ组分别为９７０％、９４１％、１０００％、９７２％。Ｘ线病灶有效率依序为９６０％、９７６％、１０００％和９４４％。５个月组与６个月组空洞关闭率分别为７７％及７６％。各组相互比较均无显著性差异（Ｐ＞０．０５），未见严重副作用。（２）３０５例完成３年随访，Ⅰ、Ⅱ、Ⅲ、Ⅳ组细菌学加Ｘ线复发分别为２、３、６和３例。结论本研究结果进一步证明Ｌ是长效、高效、安全、便于督导的新药；巩固期用Ｚ无必要；现有基本药物合理联用有可能缩短疗程为５个月，值得进一步研究。     

联合检测对菌阴肺结核诊断价值的探讨

本文探讨联合血清抗体ＰＰＤ－ＩｇＧ、ＬＡＭ－ＩｇＧ、ＴＢ－ｄｏｔ、ＩＣＴ卡及痰聚合酶链反应（ＰＣＲ）检测对菌阴肺结核的诊断价值。１对象与方法１．１选例和分组（１）实验组：菌阴肺结核７０例,３次涂片、１次培养均阴性。Ｘ线胸片有活动性肺结核征象和／或临床症状,经抗结核治疗证实为肺结核者。（２）非结核肺部疾病组共４６例。其中肺癌３２例、ＣＯＰＤ１０例、肺炎４例,均经相应检查确诊。（３）菌阳肺结核对照组４０例,其中３２例为涂（＋）培（＋）,余８例涂（－）培（＋）。１．２检测方法痰ＰＣＲ荧光法采用深圳匹基…     

Can serial qualitative polymerase chain reaction monitoring predict outcome of pulmonary tuberculosis treatment?

BACKGROUND: The aim of this study was to assess the use of qualitative one-tube nested polymerase chain reaction (PCR) for monitoring the treatment response in smear-positive pulmonary tuberculosis, and the factors determining the negative conversion of sputum smear, culture, and PCR during treatment. METHODOLOGY: A total of 53 patients receiving a standard short course of chemotherapy with 24 months follow-up period after treatment cessation were included in the study. Sputum specimens were collected serially for smear, culture, and PCR until the treatment was complete. RESULTS: The conversion rate for sputum culture, smear, and PCR at 8 weeks after treatment were 84.9, 58.5, and 47.1%, and at 16 weeks of treatment were 100, 88.7, and 79.2%, respectively. At the end of the treatment period, there were four PCR persisters, one of whom had disease relapse. Only cavitary disease had an influence over the negative conversion of the smear and PCR at 8 weeks (RR 3.5, 95% CI 1.04-11.95, P=0.04 for smear; RR 5.06, 95% CI 1.196-21.42, P=0.03 for PCR). CONCLUSION: Qualitative PCR was not useful for monitoring therapy in smear-positive pulmonary tuberculosis. Mycobacterium DNA was cleared slowly in cavitary disease. The PCR may be performed at the time of treatment cessation to identify those with potential for disease relapse.     

短程化疗结束后痰菌阴性肺结核患者复发情况随访观察

目的：评价WHO西太区结核病控制示范区项目（江苏）的控制效果，分析患的有关复发因素。方法：对西太区项目发现、登记的涂阳肺结核患，在治愈后的2－3年进行痰结核分支杆菌厚涂片检查，观察细菌学复发情况；对涂阳患治疗前的排菌情况和痰菌转阴时间与复发的关系，进行历史性队列研究。结果：随访观察的1730例治毹涂阳肺结核患，经痰涂片检查，痰菌复阳57例，复发率为3．29％。治疗后第2个月、第3个月和大于3个月转阴的初治涂阳患复发率分别是2．1％、5．3％和8．8％（P＜0．01）。治疗前不同排菌量患的复发率有一定差异，但差异无显性（P＞0．05）。结论：WHO西太区结核病控制示范区项目（江苏）的远期治疗效果是满意的。治疗后第3个月及大于3个月开始痰菌转阴的初治涂阳患复发率明显高于第2个月转阴的患。     

Influence of pre-treatment bacillary load on treatment outcome of pulmonary tuberculosis patients receiving DOTS under revised national tuberculosis control programme

OBJECTIVE: To study the influence of initial bacillary load on sputum conversion rates and treatment outcome of new smear positive pulmonary tuberculosis patients. METHODS: A retrospective study was done among 2938 new smear positive pulmonary tuberculosis patients, registered at the peripheral centres, covering a population of 1.6 million in Delhi, India. The patients pre-treatment sputum smears were graded as 1+, 2+ or 3+ based on three samples. Patients were given intermittent short-course chemotherapy under supervision and the treatment outcome was analysed. RESULTS: Sputum conversion rates among patients graded as sputum 3+ and rest of the patients (combined graded sputum 1+ and 2+) at the end of two months were 62.2% and 76.8% respectively (p<0.001), and at the end of three months were 81.3% and 89.5% respectively (p<0.001). Cure rates among same group of patients were 76.6% and 85.1% respectively (p<0.001), and failure rates were 7.7% and 4.5% respectively (p<0.001). CONCLUSIONS: Under field conditions even with directly observed treatment (DOT) new smear positive patients with heavy bacillary load showed statistically significant poor sputum conversion rates at two and three months and higher failure rates as compared to patients with lesser bacillary load. To investigate possible reasons for this poor response and possible solutions further studies are needed.     

TaqMan聚合酶链反应技术检测结核分支杆菌DNA及其临？…

目的 探讨ＴａｑＭａｎ聚合酶链反应（ＴａｑＭａｎ－ＰＣＲ）技术在肺结构诊断中的价值。方法 对１６８例活动性肺结核、５７例肺癌患者的痰和外周血及３４－例健康对照外周血,同时应用ＴａｑＭａｎ－ＰＣＲ、ＰＣＲ检测,并与痰涂片法、ＢＡＣＴＥＣ法及改良罗氏培养法结果进行比较。结果 ＴａｑＭａｎ－ＰＣＲ检测痰和外周血总的阳性率分别为５３．０％和６１．３％,显著高于ＰＣＲ、痰涂片法、ＢＡＴＣＴＥＣ法及改良罗氏培     

母牛分支杆菌菌苗在初治肺结核治疗中的作用

目的 观察和评价母牛分支杆菌菌苗（微卡菌苗）在初治肺结核免疫治疗中的疗效及安全性。方法 采用随机配对分组法将342例初治菌阳肺结核患者分入微卡菌苗治疗组（M组,171例）和对照组（C组,171例）。M组化疗方案为2HRZE／2HR,C组为2HRZE／4HR。M组加用微卡菌苗治疗6个月,C组不用微卡菌苗。结果 M组第1个月涂片阴转率36．8％,培养转率19．3％;第2个月涂片阴转率80．1％,培养阴转率85．9％。C组第1个月涂片阴转率19．9％,培养阴转率19．3％;第2个月涂片阴转率54．4％,培养阴转率67．8％。头2个月痰菌阴转率M组显著高于C组（P＜0．01）。疗程满6个月后M组涂片阴转率98．2％,培养阴转率99．4％;C组涂片阴转率98．8％,培养阴转率98．8％。M组与C组治疗6个月痰菌阴转率无显著性差异（P＞0．05）。病灶吸收好转及空洞缩小关闭速度,M组优于C组。M组的细胞免疫功能显著改善。1年后随访M组和C组的的细菌学复发率分别是3．0％和5．6％（P＞0．05）。结论 微卡菌苗能改善初治肺结核患者的细胞免疫功能,加快痰菌阴转、病灶吸收及空洞缩小关闭的速度,缩短短程化疗疗程。不良反应少且较轻微。复发率底。微卡菌苗可用作初治肺结核的免疫治疗和短化的辅助治疗。     

Survival and relapse rate of tuberculosis patients who successfully completed treatment in Vietnam.

SETTING: Reported tuberculosis (TB) cure rates are high in Vietnam with the 8-month short-course chemotherapy regimen. However, long-term treatment outcomes are unknown. OBJECTIVE: To assess survival and relapse rates among patients successfully treated for new smear-positive pulmonary tuberculosis (PTB). METHODS: A cohort of patients treated in 32 randomly selected districts in northern Vietnam were followed up 12-24 months after reported cure or treatment success for survival and bacteriologically confirmed relapse. Measurements included sputum smear examination, culture and interview for recent treatment history. RESULTS: Of 304 patients included in the study, no information was available for 31 (10%) and 19 (6%) had died. Bacteriology results were available for 244 (80%). The median interval between treatment completion and follow-up was 19 months. Relapse was recorded in 21/244 (8.6%, 95%CI 5.4-13), including 9 (4%) with positive sputum smears, 3 (1%) with negative smears but positive culture and 9 (4%) who had started TB retreatment. Four of 12 culture-positive relapse cases (33%) had multidrug-resistant strains. If the definition of relapse was extended to include death, reportedly due to TB, the relapse proportion was 26/263 (9.9%, 95%CI 6.6-14). CONCLUSION: A substantial proportion of patients (15%) had died or relapsed after being successfully treated for TB in northern Vietnam.     

A case of multidrug-resistant pulmonary tuberculosis cured by the regimen including thiacetazone

A 30 years-old-male was referred to our hospital for surgical treatment of multidrug-resistant tuberculosis in April 1998, three years after diagnosis of tuberculosis. All first-line anti-tuberculosis drugs and second-line anti-tuberculosis drugs were resistant on drug susceptibility tests by Ogawa medium. The right upper lobectomy was done because of massive hemoptysis and enlargement of cavitary lesion in June 1998, but this surgical operation was complicated with, bronchial fistula and chronic empyema. Open drainage surgical treatment for chronic empyema was done one month after lobectomy. Sputum culture for M. tuberculosis converted 4 months after the lobectomy, but bacteriological relapse occurred 17 months after initial operation. The new cavitary lesion on middle left lung field developed and sputum smear and culture were continuously positive. Immunotherapy with interferon-gamma via aerosol didn't show any clinical effect. Thiacetazone, sparfloxcin, pyrazinamide, cycloserine was prescribed after 21 months of the initial operation. Four months after changing the regimen sputum smear and culture converted to negative. Chemotherapy was terminated in June 2003, two years after negative conversion. Three years after the termination of treatment no relapse occurred. We considered thiacetazone was effective in this case, because all of the drugs was companied with thiacetazone were resistant by the drug susceptibility tests and were previously used.     

Nine month chemotherapy with INH and rifampicin for non-cavitary pulmonary tuberculosis

In order to know the adequate duration of the chemotherapy with two drugs (INH + RFP) for pulmonary tuberculosis with non-cavitary minimal radiological findings (minimal case), 278 cases with minimal lesion which had completed 9 months' chemotherapy, were observed for more than six months up to 5 years (mean duration = 54.4 months). Of them, 60 cases were bacteriologically confirmed by smear and/or culture examination. Many cases showed further improvement in radiological findings even after the end of the chemotherapy. Of 180 cases of initially infiltrative type (GAKKEN B type), 10 cases showed the enlargement of shadow radiologically, but were not regarded as relapsed cases, because they remained bacteriologically negative and the shadow improved in 1-2 months without additional chemotherapy. Only 3 cases (1.1%) were regarded as relapsed cases because of the positive bacteriological conversion and aggravation of the shadow. They were initially sputum negative. It can be concluded that for radiological minimal cases, nine months is enough for the duration of chemotherapy when the INH-RFP regimen is used.     

空洞清除术治疗复治菌阳空洞型肺结核临床研究

目的　寻找通过外科手段治疗内科无法根治的复治耐多药菌阳重症空洞型肺结核的有效途径。方法　１９８１ ～１９９８ 年６ 月采用空洞清除术治疗复治耐药空洞型肺结核１０４ 例。切除覆盖在空洞外侧的肋骨后段，游离相应部位的带蒂肋间组织，剪除空洞外侧壁，彻底清除空洞内容物，刮出新鲜创面，有机酸杀灭可能残留的结核分支杆菌，将准备好的肋间肌瓣填充于空洞残腔内，缝合固定。结果　１０４ 例中出院时治愈１０３ 例（９９０ ％ ） ，其中一次手术治愈１０１ 例（９８１ ％ ） ，好转１ 例（１０ ％ ） 。随访９８ 例中，复工率９６ ％ ，无手术死亡和复发者。结论　空洞清除术设计合理，是治疗内科无法根治的复治菌阳空洞型肺结核的有效方法。