Table of Content

10 October 2009, Volume 31 Issue 10

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Expression and immunological characteristics of recombinant fusion protein 38kD-16kD of Mycobacterium tuberculosis

Liu Zhonghua, Ding Yuansheng，Bi Aixiao，Feng Yonghong, Jin Ruiliang,Yang Hua, Qin Linhua，Hu Zhongyi

Chinese Journal of Antituberculosis. 2009, 31(10):  565-568. 

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ObjectiveTo clone, express and purify the recombinant fusion protein 38kD-16kD of Mycobacterium tuberculosis, and to evaluate its potential value in TB serodiagnosis. MethodsThe genes coding 16kD and 38kD proteins were amplified by polymerase chain reaction (PCR) from the genome of Mycobacterium tuberculosis H₃₇Rv,and then cloned into expression vector pET21a,respectively. The fusion proteins 38kD-16kD were expressed in E.coli BL21(DE3). It was purified by his-tag of the fusion protein and renaturated by semipermeable membrane. Recombinant protein was analyzed by Western-blot. The purified recombinant protein was used as antigens to screen the sera from the patients with pulmonary TB (n=96),as well as other pulmonary disease (n=50), and clinically healthy controls(n=38) by ELISA. ResultsThe recombinant plasmid pET21a-38kD-16kD was obtained. The relative molecular mass of the protein was about 65000 Da by SDS-PAGE analysis. Specific immunogenicity of the recombinant protein was confirmed by western blot analysis. The Results of ELISA showed that the sensitivity and specificity of 38kD-16kD antigen were 80.2%(77/96) and 90.9%(80/88),respectively. ConclusionThe recombinant fusion protein 38kD-16kD was successfully expressed and purified, and may be a potential candidate of serodiagnostic reagent.

The analysis of CD4 CD25 regulatory T cells and Foxp3 mRNA in the peripheral blood from the patients with tuberculous pleurisy

Lv Wei¹, Xue Keying², Wang Chengguo², Zhang Hongwei³

Chinese Journal of Antituberculosis. 2009, 31(10):  569-572. 

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ObjectiveTo investigate the changes of CD4 CD25 regulatory T cells (CD4 CD25 Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from the patients with tuberculous pleurisy, to elucidate the possible roles of CD4 CD25 Treg in the development of tuberculous pleurisy. MethodsThe peripheral blood samples were collected from 30 healthy controls (normal control group) and 32 patients with tuberculous pleurisy. The CD4 CD25 Treg ratio and Foxp3 mRNA in PBMCs were detected By flow cytometry and RT-PCR. ResultsThe CD4 CD25 Treg ratio and Foxp3 mRNA in PBMCs from tuberculous pleurisy group were higher than that from normal control group (P<0.05). ConclusionThe increase of CD4 CD25 Treg ratio and its function in PBMCs may be responsible for pathogenesis of tuberculous pleurisy.

Identification of strains isolated from tuberculosis outbreak in a high school

Wang Shengfen¹, Wang Li², Yang Lijun²,Jiang Guanglu¹, Xia Hui¹, Zhou Yang¹, Shang Mei, Li Qiang¹, Zhao Bing¹, Zhao Yanlin¹

Chinese Journal of Antituberculosis. 2009, 31(10):  573-578. 

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ObjectiveTo identify the bacterial species and strain types of isolates from tuberculosis outbreak in a high school. MethodsFrom May to August 2006, one teacher and 64 students developed clinical symptoms of active tuberculosis in a high school, Liaoning Province. Clinical isolates from sputa were collected and identified by phenotypic tests; conventional biochemical methods; amplification of hupB gene, dnaA-dnaN region, NTF-1 region, Spoligotyping and micobacterial interspersed repetitive unit (MIRU) typing; partial sequencing of 16S rRNA, 16S-23S internal transcribed spacer (ITS) region, hsp65 gene (441 bp) and PCR restriction enzyme analysis of the hsp65 gene. ResultsClinical isolates were identified as Mycobacterium tuberculosis complex and non-tuberculous mycobacteria (NTM) by conventional biochemical methods. Subsequent analyzed by phenotypic tests and amplification of hupB gene, dnaA-dnaN region, NTF-1 region, Spoligotyping and MIRU genotyping, these Results indicated that the species identification of mycobacterium tuberculosis complex as mycobacterium tuberculosis Beijing genotype modern strain, the genotype profile of MIRU was 223325173533. By partial sequencing of 16S rRNA, 16S-23S ITS region, hsp65 gene (441 bp) and PCR restriction enzyme analysis of the hsp65 gene supported the species identification of MOTT as M. porcinum. ConclusionThe mycobacterium species isolated from patients involved in tuberculosis outbreak are mycobacterium tuberculosis Beijing genotype modern strain and M. porcinum, the significance of M.porcinum in tuberculosis outbreak should be further studied.

Diagnostic value of combination with pleural biopsy and Mycobacteria Culture from pleural effussion in Tuberculous Pleurisy

Lin Minfang¹, Fang Sufang², Huang Hengcan²

Chinese Journal of Antituberculosis. 2009, 31(10):  579-582. 

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ObjectiveTo explore the diagnostic value of combination with mycobacteria culture from pleural effusion and pleural biopsy or alone in patients with tuberculous pleurisy.Methods Mycobacteria culture from pleural effusion and pleural biopsy were carried out in 270 patients(including 141 cases with pulmonary tuberculosis) with tuberculous pleurisy. We observed and analyzed the positive rates of culture, biopsy and the combination.ResultsThe positive rates of Mycobacteria culture from pleural effusion, pleural biopsy, and combination with two Methods in patients with tuberculous pleurisy were 41.1%, 62.5% and 80.0% respectively. The rate of combination with two method were significant higher than those of mycobacteria culture from pleural effusion(χ²=85.476，P<0.01) and pleural biopsy (χ²=14.892，P<0.01).ConclusionThe Methods of mycobacteria culture and pleural biopsy are safe and accurate with high value to dianose tuberculous pleurisy. Combination with pleural biopsy and mycobacteria culture may improve the rate of diagnosis. In addition, mycobacteria culture can identify whether mycobacteria strain isolated from pleural effusion is multidrug-resistant tuberculosis(MDR-TB) or non-tuberculous mycobecteria(NTM).

Analysis of the diagnosis and treatment situation of tuberculosis patients reported on internet before referral in place in non-tuberculosis control institutions in Anhui Province

Wang Aimin¹，Wang Jixiang¹，Xu Yulei²，Zhang Daomin³，Zhang Shujun⁴，Lu Lunshan⁵

Chinese Journal of Antituberculosis. 2009, 31(10):  583-585. 

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ObjectiveTo investigate the situation of diagnosis and treatment and referral of tuberculosis patients in non-tuberculosis control institutions in Anhui Province, and to provide scientific evidence for the development of future referral policies. MethodsAccording to the geographic structure, Xiao county，Dingyuan county，Feixi county and Guichi district were chosen as the study site for those place can basically reflect the situation in Anhui Province. A questionnaire survey of the situation of diagnosis and treatment of tuberculosis patients reported on internet before referral in place was conducted to the tuberculosis patients who were reported on internet and treated in special TB clinic during April 1 to September 30, 2008. Results83.4% of tuberculosis patients preferred a general hospital and/or township health centers when they had symptoms. 90.8% was referred. The patients treated in other medical facilities had spent 1422 Yuan per capita. Only 14.9% of patients had accepted the sputum smears test. 69.7% had patient delay. The annual income was about 5056 Yuan and 2479 Yuan per capita before and after they got sick. ConclusionsThe situation of referral of TB patients in non-tuberculosis prevention and control institutions is good. However, patients spent so much during the treatment and a larger decline in income happens after the sick, which may affect the further treatment compliance.

Pilot study on practicing TB control program in provincial TB special hospital in Shandong Province

Geng Hong, Liu Zhimin, Zheng Jianli, Li Haitao,Bi Xiuli, Wang Xuedong, Wang Xiuting

Chinese Journal of Antituberculosis. 2009, 31(10):  586-588. 

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ObjectiveTo evaluate the implementation effect of TB control program pilot in Shandong TB special hospital. MethodsThe statistical and monitoring data from July 2007 to August 2008 of the hospital were colleted and analyzed. ResultsThe registration rate, reporting rate and referral rate of tuberculosis suspects and patients increased month by month, the registration rate and reporting rate increased from 70% before pilot to 100%, referral rate increased from less than 10% to 94.9%, the referral arrival rate increased from 5% to 44.9%. The expected targets of registration, reporting and referral were achieved. ConclusionsSpecial hospital is the main place for TB patients consulting, it is imperative to carry out TB control program in hospital and establish mechanisms of cooperation with tuberculosis prevention and control institutions, which play a positive role in improving case detection and ensuring the cure rate.

Analysis on referral and tracing of tuberculosis cases reported from network by non-TB control institutions in 2005—2008 in Guangdong

Fang Lanjun, Zhong Qiu, Zhou Lin, Li Jianwei, Pan Qili

Chinese Journal of Antituberculosis. 2009, 31(10):  589-592. 

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ObjectiveTo analyze the status of referral and tracing of tuberculosis cases reported on internet by non-TB control facilities in 2005—2008, and to explore measures for improving tuberculosis cases finding. MethodsThe monthly and quarterly reports of TB control from 2005 to 2008 in Guangdong were collected and analyzed. ResultsThe referral arrival rate, tracking arrival rate and overall arrival rate in 2005—2008 was 42.5%, 39.0% and 64.9% respectively. 52.8% of the cases who were reported and transmitted directly through the internet by non-TB control institutions were diagnosed as TB. The referral arrival rates of bacteriological positive cases、bacteriological negative cases and no sputum smear cases reported from internet-based reporting system were 66.9%、58.0%and 37.9% respectively, the diagnosis rates of bacteriological positive cases、bacteriological negative cases and no sputum smear cases traced in place by TB control institutions were 97.6%、79.8% and 43.6% respectively. ConclusionsNon-TB control institutions play an important role in TB case detection. The case detection of pulmonary tuberculosis will be improved by establishment of standard registration, reporting and referral system, which will increase the sputum examination rate of patients reported through internet-based reporting system.

Analysis of the status of TB infection in HIV/AIDS patients in Hubei Province

Zhao Dingyuan, Ye Jianjun, Xiong Changfu, Yang Chengfeng, Zhou Liping, Luo Junmin, Wang Xiaojing, Li Aiguo

Chinese Journal of Antituberculosis. 2009, 31(10):  593-595. 

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ObjectiveTo find out the status of TB infection among people infected with HIV/ADIS patients and to obtain baseline data to prevent HIV/TB co-infection. MethodsQuestionnaire together with CD4, PPD tests, X-ray and sputum microscopy was applied to HIV/AIDS patients available for follow-up visits in 11 counties/districts. ResultsTB prevalence rate of the 622 patients visited was 3.9%, PPD positive rate of HIV/TB co-infection patients was 54.2%. The PPD positive rate was 19.0% in patients visited. The PPD positive rate increased with age. ConclusionsThe prevalence rate of tuberculosis in HIV/AIDS patients is higher than that in general population, while PPD positive rate is lower than that in general population. PPD test result of HIV/AIDS patients should only be the reference index for diagnosis of active TB.

Clinical analysis of early respiratory failure after operation of pulmonary tuberculosis

Han Ming,Zhang Jing, Wang Zidan, Chan Xiaojia

Chinese Journal of Antituberculosis. 2009, 31(10):  597-600. 

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ObjectiveTo evaluate the morbidity, mortality and risk factors of early postoperative respiratory failure after lung resection of pulmonary tuberculosis. Methods302 patients with pulmonary tuberculosis performed by surgical resection of lung were recruited from Jan 2002 to Dec 2007 in our hospital and analyzed retrospectively. Data from observation group included 19 patients with early respiratory failure after operation and a control group included 35 patients sampled radomly from 283 patients with non-failure of respiratory were observed and carried out by univariate analysis. Multivariate analysis were carried out further for data with significant difference by univariate analysis to identify risk factors associated with early respiratory failure followed by lung resection of pulmonary tuberculosis. Hospital days and mortality were compared between the respiratory failure group and non-failure of respiratory group. ResultsAmong 302 cases, 19 cases (6.3%) were diagnosed as respiratory failure. Logistic regression analysis showed that age and operative Methods were associated with early postoperative respiratory failure. The mortality and hospital days were significantly different between the respiratory failure group and non-failure of respiratory group. Hospital costs was increasing significantly. ConclusionsPatients with older age and extensive surgical range have a higher risk of respiratory failure after lung resection of pulmonary tuberculosis. Early postoperative respiratory failure does not only increase risk of mortality of patients with pulmonary tuberculosis, but to prolong hospital days and to increase hospital costs.

Clinical experience in diagnosis and treatment of 70 cases with intestinal TB

Du Debing, Yu Ping

Chinese Journal of Antituberculosis. 2009, 31(10):  602-604. 

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ObjectiveTo investigate the clinical characteristics and reasonable surgical treatment of intestinal tuberculosis. MethodsDiagnosis and treatment in 70 cases with intestinal tuberculosis hospitalized from 2000 to 2007.were analyzed retrospectively. ResultsThe longest time from the initial onset to confirmed diagnosis was 25 months and the shortest time was 3 months. The average time was 3 months. Of 70 cases, 11 cases received antituberculosis drugs and surgical treatment, in which 10 cased were cured except one case died due to without regular antituberculosis therapy. Other cases without surgical operation were cured after receiving regular antituberculosis therapy and supportive care for one year. ConclusionEarly diagnosis, timely regular antituerculosis therapy and surgical operation with strict operative indication are the key to cure the disease.

Clinical study on complete debridement in chest wall tuberculosis

Xie Jichen, Sun Liuan, Ma Yajie, Wei Zhixian Wang Zhangen,Xiao Xiuting

Chinese Journal of Antituberculosis. 2009, 31(10):  605-607. 

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ObjectiveTo evaluate the clinical efficacy of complete debridement in chest wall tuberculosis. MethodsWe analyzed 168 patients with chest wall tuberculosis hospitalized in the fifth people’s hospital of Puyang from September 2001 to January 2008. Of 168 patients, 90 patients received complete debridement and 78 patients received traditional focal debridement. Operation time, postoperative hospital days and postoperative complications were compared between two groups. ResultsIn the complete debridement group, the operation time and postoperative hospital days were 75.00±28.52 minutes and 12.66±4.46 day respectively. There were 4 patients with subcutaneous effusion at operation incision, 3 with delayed healing because of incision infection, no patient without healing, and 2 with relapse at original incision in the complete debridement group. However, the operation time and postoperative hospital days were 72.31±25.20 minutes and 14.16±5.41 day respectively in traditional operation group. And there were 12 patients with subcutaneous effusion at incision and 9 with delayed healing due to incision infection and 4 with unhealed incision and 1 with relapse at original incision. ConclusionsThe rate of postoperative complications and postoperatvie relapse of patients with complete debridement are lower than those of patients with traditional operation. Complete debridement is value to be applied for the clinic.