Surgical Management of Mitral Valve Infective Endocarditis with Accidental Finding of Open Pulmonary Tuberculosis

Objective: This case report aimed to highlight intersections of TB and Cardiovasular diseases which carry high morbidity and mortality rates. Methods: We are reporting the surgical management of forty seven years female who had back ground history of IDDM (Insulin dependent diabetic Mellitus), ESRD (End stage renal disease) on HD (haemodialysis) also she had left subclavian artery stenosis, and paroxysmal atrial fibrillation. She was diagnosed with mitral valve infective endocarditis and found accidently to have an open pulmonary tuberculosis (TB) on the day before surgery. Results: She was started on first line anti-TB treatment. She was isolated in her private room and airborne precautions measures applied. The patient underwent a tissue Mitral Valve replacement and tricuspid valve repair annuloplasty. Special precautions were applied in Theatre and on cardiopulmonary bypass Machine guided by KAMC-J disinfection protocol. The patient made good recovery postoperatively. She was discharged well on day 7 post operatively. Conclusion: Intersections of TB and cardiovasular diseases carry high morbidity and mortality rates. Early diagnosis and early anti tuberculosis treatment can surely improve the patient prognosis. Our decontamination and disinfective procedures are recommended. Cases like this should be monitored long term for the development of further cardiovascular complication.

Natural Remedies against Multi-Drug Resistant <i>Mycobacterium tuberculosis</i>

Tuberculosis (TB), caused by Mycobacterium tuberculosis is an infectious deadly disease and the treatment of which is one of the most severe challenges at the global level. Currently more than 20 chemical medications are described for the treatment of TB. Regardless of availability of several drugs to treat TB, the causative agent, M. tuberculosis is nowadays getting resistant toward the conventional drugs and leading to conditions known as Multidrug-resistant tuberculosis (MDR-TB) and extensively drug resistant tuberculosis (XDR-TB). This situation has terrified the global health community and raised a demand for new anti-tuberculosis drugs. Medicinal plants have been used to cure different common as well as lethal diseases by ancient civilizations due to its virtue of variety of chemical compounds which may have some important remedial properties. The aim of the present review is to focus the anti-tubercular medicinal plants native to India as well as the plants effective against MDR or XDR-TB across the globe. In the present review, we have addressed 25 medicinal plants for TB and 16 plants effective against MDR-TB testified from India and 23 herbal plants described for MDR-TB across the world during 2011-2015. These herbal plants can serve as promising candidates for developing novel medications to combat multidrug resistant M. tuberculosis.

Identification, Synthesis, Isolation and Spectral Characterization of Multidrug-Resistant Tuberculosis (MDR-TB) Related Substances

Several related substances were detected at trace level in (2R)-2,3-dihydro-2-methyl-6-nitro-2-[[4-[4-[4-(trifluoromethoxy)phenoxy]-1-piperidinyl] phenoxy] methyl]imidazo[2, 1-b]oxazole drug substance by a newly developed high-performance liquid chromatography method. All related substances were characterized rapidly but some impurities were found to be intermediates. Proposed structures were further confirmed by characterization using NMR, FT-IR, and HRMS techniques. Based on the spectroscopic data;unknown related sub-stances were characterized as 1-(Methylsulfonyl)-4-[4-(trifluoromethoxy) phenoxy]piperidine;4-{4-[4-(Tri-fluoromethoxy)-phenoxy]piperidin-1-yl}phenol and 4-{4-[4-(trifluoromethoxy)phenoxy]piperidin-1-yl}phenyl methane sulfonate;4-Bromophenyl methane sulfonate, Ethyl 3,6-dihydro-1(2H)-pyridine carboxylate, (2S)-3-(4-Bromophenoxy)-2-hydroxy-2-methylpropyl methane sulfonate, (2S)-3-(4-Bromophenoxy)-2-methylpropane-1,2-diyldimethane-sulfonate, (2S)-2-Methyl-3-(4-{4-[4-(trifluoromethoxy) phenoxy]-piperidin-1-yl} phenoxy)-propane-1,2-diyldimethane sulfonate, (S)-3-(4-Bromophenoxy)-2-methyl-propane-1,2-diol and corresponding Enantiomer, (2R)-2-[(4-Bromo-phenoxy)methyl]-2-methyloxirane and (2R)-2-[(4-bromophenoxy)methyl]-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b][1,3]oxazole. A possible mechanism for the formation of these related substances is also proposed.

First Line Anti-Tuberculosis Drugs Resistance Patterns of <i>Mycobacterium tuberculosis</i>Isolates from Newly Diagnosed Cases of Tuberculosis

Introduction: Tuberculosis is a major cause of mortality and morbidity world-wide. Anti-tuberculosis drugs have been used for many decades but resistance to them is now widespread. Globally 5% of tuberculosis cases and in India 3% among new TB cases. This study was planned to know the pattern of first line anti-tuberculosis drug resistance in south Gujarat, Surat region in newly diagnosed patients of tuberculosis. Material and Methods: 350 samples were processed for homogenisation and concentration using 4% NAOH-2.9% trisodium citrate. Processed samples were inoculated in liquid medium that is MGIT (Mycobacterial growth indicator tube). Positive samples for M. tbwere processed further for first line anti-tuberculosis drugs sensitivity testing (DST). Reading was taken by using MicroMGIT system. Result: Out of 350 samples 59 (17%) were positive samples, of which 48 (13%) were M. tb and 11 (3%) were non tuberculous mycobacteria. Out of 48 samples 2% (1 isolate) was resistant to isoniazid and Rifampicin while 2% were monoresistant to isoniazide, 2% monoresistant to streptomycin. No rifampicin monoresistant was detected. Conclusion: Such study may help in control of tuberculosis at regional and national level which would in turn help in planning of measures to control Multi-drug resistance tuberculosis. Continuous surveillance should be applied to know the periodic changing patterns and trend in Drug resistant tuberculosis.

Synergistic Effects of 18β-glycyrrhetinic Acid Combined with Antituberculosis Drugs against Mycobacterium tuberculosis

The in vitro antibacterial activities of 18β-glycyrrhetinic acid alone or combined with first-line antituberculosis drugs including isoniazid（INH）,rifampicin（RFP） and streptomycin（SM） against Mycobacterium tuberculosis were detected using MABA method.The minimum inhibitory concentrations（MICs） of18β-glycyrrhetinic acid against M.tuberculosis H37Rv（ATCC 27294） and M.bovis（ATCC 19210） were 50 and 100 μg/m L,respectively.The MICs of two clinical drug-susceptible isolates and six drug-resistant isolates were 25-50 and 100-200 μg/m L,respectively.As 18β-glycyrrhetinic acid combined with INH,RFP and SM,they exhibited synergistic effects against six drug-resistant isolates,and MICs decreased significantly：MIC of INH decreased by 2-32 folds（FICIs 0.125-0.375）;MIC of RFP decreased by 4-8 folds（FICIs 0.240-0.490）;MIC of SM decreased by 4-16 folds（FICIs 0.165-0.460）.Traditional medicine monomer had low cytotoxicity on normal cell BHK-21 and could restraint SMMC fission.The results showed that 18β-glycyrrhetinic acid combined with anti-TB drugs（INH,RFP and SM） had good antibacterial activity against M.tuberculosis.These findings indicated that 18β-glycyrrhetinic acid might serve as the potential therapeutic compound for future development of anti-TB drugs.

Fast and Accurate Identification of <i>M. tuberculosis</i>Complex Using an Immunochromatographic MPT64 Antigen Detection Test

Background: A new rapid Immunochromatographic test (ICT) kit (MPT64 TB Ag Kit) for detection of MPT64 Antigen in M. tuberculosis (MTB) isolates used for rapid identification of MTB isolates developed by SD (Standard Diagnostics) Bio line, South Korea was evaluated. The ICT is a rapid, reliable and cheaper method that can be used instead of conventional biochemical tests for confirming MTB in culture isolates in resource limited laboratories. The study also evaluated the ability of ICT to detect MPT64-Antigen before the micro MGIT could signal positive. Material/Methods: A total of 450 sputum samples of individual patients were used for the study. 152 isolates of Mycobacteria were recovered from solid and liquid media. These strains were tested for the detection of MPT64-antigen. H37Rv strain was served as the positive reference control and also used for early detection of Antigen experiment. Findings: The development of bands on both test and sample region when H37Rv strain was tested were seen (MPT64 antigen positive). When 138 MTB isolates were tested, it showed a similar banding pattern indicating 100% sensitivity. MPT64 band formation was not detected in any of the 14 isolates indicating 100% specificity. Both PPV & NPV were 100%. All the isolates negative for MPT64 Ag were confirmed as MOTT by conventional bio-chemical PNBA. The H37Rv strain showed a faint band from the 2nd day onwards from inoculation till 3rd day in the earlier Antigen detection experiment. Conclusion: Rapid identification of MTB culture isolate is a pressing need for diagnosis and proceeding to perform drug susceptibility testing. MPT64 TB Ag detection ICT kit is a rapid, reliable method, good substitute for molecular identification methods, and conventional biochemical test which is time-consuming and technically demanding. The early detection of Antigen can be used as an effective tool in diagnosis.

结核分枝杆菌Mtb8.4DNA疫苗构建及免疫保护研究

目的评价构建结核菌DNA疫苗pVS84免疫小鼠体外产生细胞因子的能力和抵抗结核分枝杆菌H37Rv攻击的效果。方法利用基因技术将结核菌Mtb8.4插入pVAXl载体,构建表达结核菌Mtb8.4蛋白(分泌型)的DNA疫苗pVS84。将雌性C57BL/6小鼠分成5组,每组20只,分别用pVS84、pVAXl、plL2S和PBS分别各免疫3次,每次均间隔2周,以等剂量加强免疫2次。另一组用BCG(105CFU)皮内注射免疫1次。每组10只鼠在最后一次加强免疫后,无菌取脾培养,检测上清细胞因子。另10只鼠则用结核菌H37Rv经静脉攻击,2周后取脾、肝和肺培养结核菌并进行菌落计数。结果pVS84免疫组鼠脾淋巴细胞培养上清的mIL-2和mlFN-Y平均含量分别为(290.0±112.9)pg/ml和(501.7±79.4)pg/ml,显著高于3个阴性对照组(P<0.001),与BCG免疫组无显著性差异(P>0.05)。5个组的平均mIL-6和mIL-10无显著性差异。pVS84免疫组小鼠的脾、肝和肺的平均结核菌载量分别为(39176.9±3702.6)、(18780.1±2535.8)和(24410.3±1846.8)CFU/g,分别低于pVAXl、plL2S和PBS三个阴性对照组的相应器官的结核菌载量(P<0.001),仅脾结核菌载量显著高于BCG免疫对照组。结论构建的DNA疫苗pVS84能够刺激机体产生抗结核菌所需的Thl型免疫应答,免疫鼠抵抗H37Rv攻击的能力与BCG接近。

Investigation of tuberculosis clusters in Dehradun city of India

Objective:To investigate the presence of statistically significant geographical clusters of tuberculosis(TB) using Geographical Information System and spatial scan statistics in Dehradun, India.Methods:The spatial scan statistic implemented with a software program,SaTScan v6.1, was used to test the presence of statistically significant spatial clusters of TB and to identify their approximate locations(P【0.05 for primary clusters and P【0.1 for secondary clusters). Geographical Information System was used for geographical analysis.Results:Significant high rate spatial clusters were identified in seven wards of the Dehradun Municipal area. Conclusions:There is sufficient evidence about the existence of statistically significant TB clusters in seven wards of Dehradun,India.The purely spatial scan statistics methodology used in this study has a potential use in surveillance of TB for detecting the true clusters of the disease.

Gene Xpert MTB/RIF 在川东某医院结核病早期诊断中的应用价值分析

目的:探讨Gene Xpert MTB/RIF技术在川东基层医疗机构中对结核病(TB)的早期诊断价值。方法:收集2020年3月至2021年2月遂宁市中心医院收治的疑似TB患者送检的标本368份,分别采用抗酸染色法、荧光定量聚合酶链式反应(PCR)法、Gene Xper t MTB/RIF技术与固体培养法对标本进行检测,比较四种检测方法的阳性检出率,并以固体培养法的检测结果为金标准,分析Gene Xpert MTB/RIF技术对结核分枝杆菌(MTB)的检测效能;以比例法药敏试验结果为金标准,分析Gene Xpert MTB/RIF技术对利福平耐药性的检测效能。结果:用抗酸染色法、荧光定量PCR法、Gene Xpert MTB/RIF技术、固体培养法对368份标本进行检测的阳性检出率分别为14.13%、34.24%、49.46%、50.82%。采用Gene Xp ert M TB/RIF技术对368份标本进行检测的阳性检出率高于荧光定量PCR法和抗酸染色法,差异有统计学意义(P<0.05)。采用固体培养法对368份标本进行检测的阳性检出率略高于Gene Xpert MTB/RIF技术,但差异无统计学意义(P>0.05)。以固体培养法的检测结果为金标准,发现采用Gene Xpert MTB/RIF技术检测疑似TB患者送检标本中MTB的敏感度为96.79%、特异度为99.45%,阳性预测值为99.45%,阴性预测值为96.77%,二者的符合率为98.10%,Kappa值为0.961。以比例法药敏试验结果为金标准,发现采用Gene Xpert MTB/RIF技术检测疑似TB患者送检标本利福平耐药性的敏感度为88.89%,特异度为99.71%,阳性预测值为96.00%,阴性预测值为99.13%,二者的符合率为98.91%,Kappa值为0.884。结论:采用Gene Xpert MTB/RIF技术检测MTB和利福平耐药性的敏感度高、特异度好且对生物安全要求较低,适合在川东生物安全防护薄弱的基层医疗机构推广使用。

Gene-XpertMTB/RIF技术在HIV合并结核分枝杆菌感染筛查中的应用

目的探讨新技术(Gene-XpertMTB/RIF,简称为Xpert)在人类免疫缺陷病毒(HIV)合并结核分枝杆菌感染筛查中的临床应用,期望为HIV/TB的感染者找到快捷、准确的检测方法或者检测方法组合。方法收集云南省传染病医院2017年3月至2018年12月收治的HIV住院患者278例痰液标本,每例患者痰液标本均用Xpert、罗氏培养法和直接涂片萋-纳抗酸染色法三种方法进行检测。结果Xpert、罗氏培养和直接涂片萋-纳抗酸染色法三种检测方法的阳性率分别为15.8%、12.6%、10.8%,Xpert与其他两种方法一致性评价,均为Kappa>0.75,一致性较好。结论Gene-XpertMTB/RIF技术是一种快速、安全、高效的分子生物学检测方法,并且能够同时检测利福平的耐药性,对艾滋病合并结核病的早期筛查具有重要意义。

4th National Anti-tuberculosis Drug Resistance Survey in Kenya

Introduction： Recently rapid development of drug resistant TB, particularly MDR TB （Multi Drug Resistant TB） and XDRTB （Extensively Drug-Resistant TB） possess a major threat to control of tuberculosis globally. Information on the extent of MDR-TB from Kenya is largely limited due to several factors. Monitoring of development of resistance is a vital tool in providing critical information for effective planning for TB control and in management of patients infected with TB. Methods： Cross-sectional with cluster design. Results： A total of 2,171 participants recruited into the study from 50 selected clusters. Prevalence of rifampicin resistance for new cases was 1.3% [95% CI, 0.8-2.0] and INH resistance was 5.5% [95% CI, 4.5-6.7]. MDR TB was found in 0.67% of new cases and 2.1% amongst previously treated TB cases. Discussion： Resistance to isoniazid in Kenya has been on the decline due to introduction of rifampicin in combined therapy. There was increase of MDR TB among new cases by 24% and decline in previously treated cases due to lethal impact of HIV. Conclusions： Although drug resistance TB is a growing problem in Kenya, resistance to isoniazid and rifampicin MDR TB is less than previously estimated. The country should continue to monitor drug resistance and ensure effective use of anti TB medicines.

Comparison of GeneXpert MTB/RIF Assay in Broncho Alveolar Lavage and Gastric Lavage Samples

Objective: To compare the yield from Gastric lavage (GL) and Broncho alveolar lavage (BAL) samples in adult patients suspected case of Tuberculosis but not producing sputum. Methodology: 80 adults with suspected case of tuberculosis but not producing sputum were recruited. 72 patients were then subjected to one gastric lavage followed by Broncho-alveolar lavage in the same morning. The collected samples were subjected to GeneXpert MTB/RIF assay. Result: Of the 72 patients samples, the mean age was 38.6 years. 41 (56.9%) were male and 31 (43.1%) were female. History of TB contact was present in 25 (34.7%) patients. 37 (51.4%) patients had GeneXpert MTB/RIF positive on BAL and/or GL samples. The GeneXpert MTB/RIF of BAL fluid was positive on 35 (48.6%), which was not significantly greater than that for specimens from GL, which was 28 (38.9%) (p > 0.05). In 26 (36.1%) cases, GeneXpert MTB/RIF was positive in both BAL and GL samples. Conclusion: This study showed the yield of GeneXpert MTB/RIF in GL was comparable to BAL to detect Mycobacterium tuberculosis complex. Patients who can’t produce sputum, GL can be a good alternative to BAL to detect Mycobacterium tuberculosis complex in resource poor areas and patients who do not tolerate Bronchoscopy.

TB AccuProbe、INNO-LIPA^(TM)RIF.TB探针和rpoβDNA测序三种方法鉴定结核分枝杆菌的研究

目的用 3种先进的技术 -TBAccuProbeDNA杂交法、INNO LIPATMRIF TB(LiPA)探针试验和rpoβDNA基因序列测定的前瞻性研究 ,以鉴定结核分枝杆菌复合物 (MTBC)和非结核分枝杆菌(NTM)及其成本效益。方法在BactecMGIT 96 0培养系统中将获得的 10 5例阳性培养物筛选出 70例MTBC和 35例NTM ,用 3种技术鉴定并检测其rpoβ基因序列突变位点。 结果TBAccuProbe杂交阳性率是 95 7% (6 7/70 ) ,LiPADNA探针阳性率是 98 6 % (6 9/70 ) ,rpoβDNA序列测定阳性率是 97 1% (6 8/70 )。 3种方法两两比较差异在统计学上均无显著意义 (P >0 0 5 )。每份阳性培养物花费时间 :TBAc cuProbe杂交法和LiPADNA探针法在 2 4小时内完成 ,全部完成需 6～ 37天 ,平均为 15天 ;rpoβ基因序列测定需 4～ 5天完成 ,全部完成需 8～ 4 0天 ,平均 17天。比传统的 4～ 8周鉴定时间减少了很多。每份标本所需费用 3种方法各为 12 0、194和 96元。在 35例NTM中 ,3种方法均为MTBC阴性 ,同时rpoβDNA序列测定鉴定了NTM的种类。结论TBAccuProbe杂交和LiPADNA探针操作简便 ,所需时间短 ,只能鉴定MTBC和NTM。但LiPADNA探针还能鉴定rpoβ基因位点突变 ;rpoβ基因序列测定所需时间稍长 ,要有特定仪器和专门人员 ,但所需费用较低 ,除能鉴定MTBC外 。

Longitudinal observation of an interferon gamma-released assay （T-SPOT.TB） for Mycobacterium tuberculosis infection in AIDS patients on highly active antiretroviral therapy

Background T-SPOT.TB is a novel test for tuberculosis infection with higher sensitivity and specificity than the traditional tuberculin skin test （TST）. However, there are no longitudinal data in the literature evaluating T-SPOT.TB for Mycobacterium tuberculosis in patients with acquired immune deficiency syndrome （AIDS） on highly active antiretroviral therapy （HAART）. The objective of this study was to assess the value of T-SPOT.TB longitudinally in AIDS patients on HAART without prophylaxis for tuberculosis.Methods A prospective observational study was conducted in 50 AIDS patients on HAART. None of the subjects had evidence of active tuberculosis. T-SPOT.TB, a T-cell-based interferon y released assay, was performed at the onset of the study and repeated 24 months thereafter. Subjects were evaluated every 6 months during the 36-month follow-up. Results Twenty-one （42%） AIDS patients on HAART tested positive by T-SPOT.TB （95% Cl 28.3%-55.7%）. The pooled spot-forming cells of early secretory antigenic target-6 （ESAT-6） and culture filtrate protein-10 （CFP-10） peptides were 68/million peripheral blood mononuclear cell （PBMC） （interquartile range 44-220）. The average number of CD4 cells in subjects was （305±152） cells/Ml and there was no significant difference in T-SPOT.TB response rates between subjects with CD4 cell counts 〈200 cells/ul （7/15 （46.7%）, 95% C/21.5%-71.9%） and those with CD4 cell counts≥200 cells/ul （14/35 （40.0%）, 95% Cl 23.8%-56.2%, P=0.662）. In the 32 subjects who completed the 24-month follow-up, 10 underwent T-SPOT.TB reversion, one had T-SPOT.TB conversion, six remained positive and 15 remained negative. None of them advanced to active tuberculosis during the 36-month follow-up.Conclusion The inactive status of tuberculosis infection may be maintained for a long period in AIDS patients on HAART.

The Role of Innate Immune Cells in the Response of Heat-Treated Mycobacterium tuberculosis (M.tb) Antigens Stimulating PBMCs

The proliferation response of γδT cells to the antigen from heat-treated Mycobacterium tuberculosis H37Ra (M.tb Ag)was used as a good model in γδT cell research.From preliminary research it is found that activated NK cells positively elevated γδT cells proliferation after simulating PBMCs with M.tb Ag.To investigate different behaviors of NK cells,γδNKT cells,γδT cells and relationships between these cell subsets,activation and proliferation of different cell subsets of PBMCs in response to M.tb Ag were analyzed.We demonstrated that NK cells,γδNKT cells and γδT cells could be activated after stimulation with M.tb Ag.γδNKT cells and γδT cells proliferated while the number of NK cells decreased after 11 day-simulation with M.tb Ag.Meanwhile,at the early time of stimulation the cytotoxicity of PBMCs was enhanced.Cellular & Molecular Immunology. 2004;1(6):467-470.

涂阳肺结核患者TB-DNA定量、γ干扰素释放试验、结核抗体检测的价值

目的探讨肺结核(TB)-DNA定量、γ干扰素释放试验(T-SPOT.TB)、结核抗体检测对活动性肺结核辅助诊断的价值。方法回顾性分析2013年7月至2014年6月入院确诊为肺结核且抗酸杆菌痰涂片阳性51例患者的TB-DNA定量、TSPOT.TB、结核抗体检测指标。结果 T-SPOT.TB检测阳性率为90.2%,灵敏度最高;其次为TB-DNA定量,阳性率为74.5%;结核抗体检测阳性率仅为37.3%,灵敏度最低。结论 T-SPOT.TB检测不受标本采集的影响,灵敏度高,对活动性肺结核的辅助诊断具有较高价值。

乙醇对结核分枝杆菌M_r 38000蛋白在大肠杆菌中可溶性表达的促进作用

目的:探讨乙醇促进结核分枝杆菌Mr38000蛋白 的可溶性表达,并获得可溶性的Mr38000重组蛋白.方法: 采用聚合酶链反应(PCR)从结核分枝杆菌H37Rv基因组中 扩增出Mr38000蛋白的编码基因,用限制性内切酶消化后插 入载体pGEX 4T 2中,将重组质粒转化大肠杆菌BL21,在乙 醇添加剂存在下,经IPTG诱导,表达GST 38融合蛋白;聚丙 烯酰胺凝胶电泳(SDS PAGE)分析融合蛋白的相对分子质量 及表达形式,免疫印迹法鉴定融合蛋白的活性.结果:经PCR 扩增后证实为Mr38000蛋白的基因,成功构建了具有正确基 因序列的重组表达载体,在大肠杆菌BL21中诱导表达融合蛋 白约占菌体总蛋白的18%.在乙醇存在下,可溶性目的蛋白的 表达量约是无乙醇时的5倍.结论:在原核表达中乙醇作为一 种添加剂可以促进结核分枝杆菌Mr38000蛋白在大肠杆菌 BL21中的可溶性表达.

腹水中TB-AB、ADA和sCD44v6的检测及意义

目的:探讨可溶性CD44v6(sCD44v6)、腺苷脱氨酶(adenosine deaminase,ADA)和结核抗体(mycobac-terium tuberculosis antibody,TB-AB)联合检测对良、恶性腹水的鉴别诊断价值。方法:取48份结核性腹水、39份恶性腹水,分别用酶联免疫吸附试验(ELISA)、酶速率法及胶体金标法检测腹水sCD44v6、ADA及TB-AB水平。结果:恶性腹水组sCD44v6水平为(102.2±30.6)ng/ml,明显高于结核性腹水组(43.5±15.3)ng/ml(P<0.01);结核性腹水组ADA水平为(55.3±22.4)U/L,明显高于恶性腹水组(21.6±12.7)U/L(P<0.01)。结核患者血清ADA水平为(44.5±17.1)U/L,明显高于恶性肿瘤患者血清ADA水平(25.6±13.8)U/L(P<0.01)。以60.5ng/ml为阳性界值,sCD44 v6诊断恶性腹水的敏感度为82.1%、特异度为87.5%。以45U/L为阳性界值,ADA诊断结核性腹水的敏感度为83.3%、特异度为87.2%。以腹水/血清ADA比值>1.0为阳性界值,诊断结核性腹水的敏感度、特异度分别为89.6%、92.3%。TB-AB诊断结核的敏感度及特异度分别为85.4%及87.2%,诊断恶性肿瘤的敏感度为5.1%。TB-AB、ADA及P-ADA/S-ADA联合检测,诊断结核性腹水的敏感度、特异度分别为95.8%、94.9%。结论:TB-AB、ADA和sCD44v6对于良、恶性腹水的鉴别诊断有重要价值。

QFT-GIT对儿童肺结核临床诊断价值的meta分析

目的系统评价QFT-GIT对儿童肺结核临床诊断价值。方法计算机检索中国生物医学数据库、中国知网(CNKI)全文数据库、维普数据库、万方全文数据库、EMBASE,PUBMED和Cochrane并提取数据,应用Stata软件进行meta分析。结果分析纳入的5个研究。结果显示,QFT-GIT作为儿童肺结核诊断试验,灵敏度点估计值为0.76,95%CI=0.65～0.84,特异度点估计值为0.98,95%CI=0.91～1.00。另外,阳性似然比点估计值为42.0,95%CI=8.5～207.8,阴性似然比点估计值为0.25,95%CI=0.17～0.37,诊断优势比OR=169,95%CI=37～775。结论 QFT-GIT具有排除诊断的能力,但鉴于受试儿童免疫状态和感染风险的差异会造成其检测能力表现存在差异,建议对该方法进行进一步评价。

结核分枝杆菌蛋白TB9.7间接ELISA方法的建立

为获得新的鉴别诊断结核杆菌感染的候选抗原,根据Gene Bank中登录的结核杆菌H37Rv的TB 9.7基因序列设计1对引物,以结核杆菌H37Rv基因组DNA为模板,PCR扩增得到TB 9.7基因DNA片段。将该扩增片段克隆于PMD18-T载体中,得到载体PMD18-T-TB9.7。分别将pET32a质粒和PMD18-T-TB 9.7质粒用限制性内切酶EcoRⅠ和XhoⅠ双酶切,并将纯化的TB 9.7基因亚克隆至pET32a中,获得重组表达质粒pET32a-TB 9.7。将其转化E.coli BL21 IPTG诱导后,经SDS-PAGE、Western blot分析鉴定,可见约30ku的外源蛋白带。表明TB9.7基因在大肠杆菌中得到了表达。用Ni-NTA Agarose试剂盒进行蛋白纯化,获得纯化的TB9.7融合蛋白,建立间接ELISA诊断方法,检测了30份结核病人的痰检阳性血清,120份健康人的血清,敏感性为56.6%,特异性为97%,与痰检结果的符合率为84%。