Table of Content

10 May 2007, Volume 29 Issue 5

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The application of protein-chip on rapid diagnosis of tuberculosis

Wang Sumin¹,Fu Yuhong,Liu Yang,et al.

. 2007, 29(5):  379-381. 

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Objective To evaluate the application value of protein-chip in detecting anti-tuberculosis(TB) antibodies in the clinical samples. Methods The serum samples from 117 patients with TB,and 103 patients with non-tuberculosis lung diseases and healthy volunteers were detected anti-TB antibodies using protein-chip.The sputum samples from 117 patients with TB were also observed acid-fast bacilli by smear and microscopy as control. Results The positive rate of antiTB antibodies were 100%(26/26),49.6%(45/91) in smear-positive and smear-negative patients with pulmonary tuberculosis by using protein chip,respectively.The negative rate of anti-TB antibodies was 98.1%(101/103) in the patients with non-tuberculosis lung diseases and healthy volunteers by detection of protein chip. Conclusion The anti-TB antibody protein-chip is a simple,rapid,high-through,sensitive,specific,low cost,and effective method for the diagnosis of TB,especially for the smear-negative TB.

Therapeutic effects of chimeric Ag85A/ESAT-6 DNA vaccines combined with chemotherapy in a mouse model infected with multi-drug resistant Mycobacterium tuberculosis

Liang Yan,Wu Xueqiong,Zhang Junxian,et al.

. 2007, 29(5):  382-385. 

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Objective To study the therapeutic effects of chimeric Ag85A/ESAT-6 DNA vaccines combined with chemotherapy in a mouse model infected with multi-drug resistant(MDR) Mycobacterium tuberculosis. Methods BALB/c mice were infected by intravenous injection in a tail vein with Mycobacterium tuberculosis clinical isolate HB240 that is resistant to Isoniazid(INH) and Rifampin(RFP) for 4 weeks,and then were randomly divided into tow groups.The mice in group 1 were treated with RFP and INH for 12 weeks.The mice in group 2 were treated by chimeric Ag85A/ESAT-6 DNA vaccines combined with INH and RFP for 12 weeks.DNA vaccines were injected intramuscularly 5 times at 3 weeks intervals.The lungs,livers and spleens were taken and observed their pathological changes,weighted and performed mycobacteria cultures at 4 or 8 weeks after terminative treatment. Results At four and eight weeks after terminative treatment,the body weights of mice in group 2 were lower than that in group 1,but it had no significant difference(P>0.05).At four weeks after terminative treatment,indexes of the lungs and spleens(0.017,0.011)from the mice in group 2 were lower than that in group 1 (0.020,0.012).No swell of spleen could be observed in 1 mouse from group 2 and 4 mice from group 1.The numbers of bacteria in the lungs and spleens from mice in group 2 decreased 2.5 times and 4 times than those in group 1,respectively.At eight weeks after terminative treatment,indexes of the lungs and livers from the mice in group 2(0.021,0.047)were lower than that in group 1(0.022,0.048)(P>0.05),but indexes of the spleens from the mice in group 2(0.008)were significantly lower than that in group 1(0.012)(P<0.05).The swell of the lymph node of lung could be observed in 1 mouse from group 2 and 3 mice from group 1,no swell of spleen could be observed in 5 mice from group 2 and 2 mice from group 1. Conclusion The therapeutic efficacy of DNA vaccine combined with chemotherapy is stronger significantly than that of chemotherapy alone in the mouse model of MDR-tuberculosis.

Rapid detection of rpoB gene mutation from Mycobacterium tuberculosis by real-time PCR and its clinical application

Chen Xiaoyou,Huang Hairong,Ma Yu,et al.

. 2007, 29(5):  386-390. 

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Objective To evaluate the value of detecting gene mutation by real-time PCR and its clinical application. Methods Targeting the most common mutation types at codon 526 and codon 531 of Rifampicin Resistance Determining Region (RRDR) of rpoB gene from Mycobacterium tuberculosis,labeled probes(526CAC, 526TAC,531TCG and 531TTG) were designed to detect 38 Rifampin-resistant clinical isolates with known RRDR sequence,24 Rifampin-sensitive clinical isolates with wild type sequence of RRDR and 5 nontuberculous mycobacteria isolates,then a method using real-time PCR was established.84 sputa from patients with pulmonary tuberculosis were analyzed by real-time PCR and drug susceptibility testing on Lowenstein-Jensen(L-J) medium.Some sputum samples were amplified by PCR with other primers and then sequenced. Results Using conventional drug susceptibility testing as control,the mutation type at codon 526 and codon 531 of rpoB gene were identified in 38 Rifampin-resistant clinical isolates,24 Rifampin-sensitive clinical isolates and 5 nontuberculous mycobacteria isolates by real-time PCR.The sensitivity and specificity were 100%,respectively.Of 84 sputum samples,62 were drug-resistant(including 48 Rifampicin-resistant samples) by conventional drug susceptibility testing,while 75 were positive by real-time PCR.Of 48 Rifampin-resistant samples,43 had TTG mutation of rpoB at codon 531,and 3 isolates had TAC mutation at codon 526 by real-time PCR.The result of sequencing showed that one mutant at codon 514 was inserted TTC,and another one had a CCG mutation at codon 511 with a GGC mutation at codon 516. Conclusion Real-time PCR is a rapid technique to detect mutation of target gene and could be an assistant tool to diagnose drug-resistant tuberculosis early.

Cloning,prokaryotic expression and purification of Mycobacterium tuberculosis Rv0867c gene

Gao Xiaopeng,Su Mingquan,Liu Jiayun,et al.

. 2007, 29(5):  391-394. 

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Objective To construct prokaryotic expression plasmid encoding Rv0867C gene from Mycobacterium tuberculosis(M.tb),to express efficiently the fusion proteins in E.coli. Methods The Rv0867c gene was amplified by polymerase chain reactions(PCR) with specific primers from genomic DNA of M.tb H37Rv strain,and cloned into plasmid pUC19.Then Rv0867C was subcloned into the expression vector pPRO-EXHT.The plasmid pPRO-EXHT-Rv0867c was transformed into E.coli DH5α and induced by IPTG.The Rv0867c fusion protein expressed was analyzed by SDS-PAGE.Recombinant His-6-fused protein was purified by Ni2+-NTA purification system. Results The length of PCR products was 1224bp.The result of DNA sequencing showed that the amplified Rv0867C gene was exactly consistent with the sequence reported in Genebank.The recombinant expressive plasmid pPRO-EXHT-Rv0867c was constructed.The E.coli DH5α strain with recombinant plasmid showed high level of Rv0867c gene expression after IPTG induction.The plasmid pPRO-EXHT-Rv0867c expressed a Rv0867c fusion protein with relative molecule mass 80,000KDa.The protein band amounted to 23.7% of total bacteria protein,SDS-PAGE analysis showed that the fusion protein mainly existed in inclusion body.The expressed protein could be purified via Ni²⁺-NTA system kit in denatured condition. Conclusion Mycobacterium tuberculosis Rv0867c gene has been cloned and expressed successfully in E.coli DH5α.The Results lay a basis for further study of fast cultivation for Mycobacterium tuberculosis.

Survey of drug-resistant M.tuberculosis in Shanghai,China

Mei Jian,Shen Xin,Shen Mei,et al.

. 2007, 29(5):  395-398. 

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Objective To determine the prevalence of primary and acquired drug resistance in M.tuberculosis,and to improve the drug resistance surveillance system in Shanghai. Methods According to WHO/IUATLD guidelines for anti-tuberculosis drug resistance surveillance,sampling method of 100% diagnostic center was adopted.All sputum culture-positive patients among local residents registered at any district(county) tuberculosis dispensary in Shanghai from February 2004 to January 2005 were enrolled.Species identification and drug susceptibility tests were performed on all samples. Results 1 076 patients were enrolled during the study period,in which 1015 patients had the Results of identification and susceptibility.Of 1 015 strains,964(95.0%)M.tuberculosis and 51(5.0%)non-tuberculosis mycobacteria were identified.The prevalence of drug-resistant M.tuberculosis among new cases and re-treatment cases was 15.4%(118/764) and 27.5%(55/200),respectively.The prevalence of MDR-tuberculosis among new cases and re-treatment cases was 3.9%(30/764)and 12.5%(25/200),respectively. Conclusion The prevalence of drug-resistant tuberculosis was severe in Shanghai,suggesting the necessary to strengthen the TB control program.

A survey of tuberculosis infection in the freshmen of universities in district Changping,Beijing

Zhao Fujun,Gao Tiejie,Liu Deshui,et al.

. 2007, 29(5):  399-401. 

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Objective To investigate the situation of tuberculin skin reaction and tuberculosis in the freshmen of universities in district Changping,Beijing,in order to adopt appropriate method and measure to control the infection source. Methods Do the tuberculin skin testing in the freshmen of universities continuously for three years in district Changping.Perform chest X-ray examination and sputum examination in tuberculin-strong-positive freshmen to find active patients with tuberculosis. Results Of 139 136 freshmen from 21 universities examined by tuberculin skin testing,the freshmen whose diameter of tuberculin skin reaction were more than 5 mm accounted for 16.48%,17.63%,and 38.06% every year for recent 3 years,respectively,in which there were 226 cases with active pulmonary tuberculosis,44 cases with tuberculous pleurisy,46 cases with inactive pulmornary tuberculosis,16 sputum smear-positive cases,3 sputum culture-positive cases. Conclusion The situation of tuberculosis in the university students was severe.The university students should be put importance to tuberculosis prevention and control,strengthen the examination at enrollment to early find and treat the patients.

The survey of increasing the rate of arrival by referring of the tuberculosis patients and suspects

Xie Yanguang,Yan Xinglu,Sun Yanbo,et al.

. 2007, 29(5):  402-405. 

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Objective To study the ways and feasibilities of further increasing the rate of arrival by referring of TB patients and suspects,so as to increase the discovery rate of patients with pulmonary TB. Methods Carrying out the some scale stratified sampling method,selecting 20 counties as project areas,in which the number of population is over 300 thousands in Heilongjiang province.Based on the available referral system,carry out a series of intervention.In the same time,select other 20 counties as control group.Study the number of TB patients and suspects who are referred and arrived. Results During the project,the arrival rate is 82.5% in the 20 project counties,and the arrival rate is 58.8% in control counties,there were significant difference between two groups(χ²=1 016.510 4,P<0.01).The number of the TB cases,the smear-positive cases and the new smear-positive cases were increased by 10.6%、11.1% and 12.7% compared with that in the same period of previous year in project counties.While in the control group,the number of the TB cases was increased by 11.2%,the number of the smear-positive and the new smear-positive patients were decreased by 1.9% and 1.3% compared with that in the same period of the previous year. Conclusion The intervention to strengthen the referral and tracing system could increase the discovery rate of patients in the area with poor baseline.

Discussion on the control and prevention of TB on community level in Anshan city,liaoning province

Chen Ligong,Li Li,Yang Fengying,et al.

. 2007, 29(5):  405-407. 

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Objective To discuss the control and prevention measures of TB patients in Anshan city,where no administrative institutions were established at the district level on TB prevention.Methods Anshan Research lnstitute for TB Control set up TB control and prevention service center,to manage the community TB prevention workers and the community TB prevention volunteers.Community TB Monitoring centers were also established on the basis of other establishments.Community TB information communication and education was also strengthened,to enforce the government prevention and control measures. Results From 1999 to 2003,280 TB prevention and control points were established,158 TB prevention workers and 493 TB prevention volunteers were mobilized. Before network reporting on the 1st of April,681 TB patients were discovered through the TB prevention network.The discovery rate on TB patients has been improved.In the meantime,the coverage area of the TB prevention and control has been increased,to make it more convenient for the TB patients from the bordering rural area.The TB knowledge has been enhanced from 71.2% in 2000 to 95.6% in 2004. Conclusion It is feasible that Anshan City took full advantage of the function of TB prevention workers and volunteers to control TB in communities.

The evaluation of midterm effect of the TB control project and programme in chen zhou

Liu songshan,Liu Jing,Cao Huifang,et al.

. 2007, 29(5):  408-411. 

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Objective To evaluate midterm effect of the implement of the TB control project and program,to provide a basis for the accomplishment of goal of program as schedule. Methods To collect and analyze the quarterly reports on TB control project from every county and other relevant materials. Results The new notification rates of smear-positive cases with pulmonary TB were 35.83/100 000 in 2001 and 52.31/100 000 in 2005,respectively.The new notification rate of smear-positive,aged TB patients in poverty counties was 73.88/100 000.The sputum negative conversion rates were 96.5% in TB patients with primary treatment and 95.1% in TB patients with re-treatment.Their cure rates were 95% and 92%,respectively.The eligible rate of sputum smear has increased by 5%.The outlay invested to the project has increased 1.68 times. Conclusion The discovery rate of new smear-positive TB cases has increased greatly.The Objective of higher cure rate and lower death rate has been reached.The DOTS and administration mode of TB control were successful in Chenzhou.

Diagnosis of pulmonary tuberculosis in general hospital

Yang Ruihong,He Quanying,Chen Ying

. 2007, 29(5):  411-414. 

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Objective To investigate the diagnosis level of pulmonary tuberculosis of in-patient in general hospital. Methods Senventy-four in-patients with pulmonary tuberculosis reported in people’s Hospital of Peking University Jan 1,2002 to May 31,2006 were retrospectively analyzed.Results Of 74 cases,71.6% were reported by respiratory department,41.9% had typical tuberculosis clinical symptom,54.1% showed that ESRs were significantly high,54.1% PPD test were positive.64.9% patients took sputum smear examination and 54.2% of them were positive.The average interval from admission to diagnosis was 13.15 days,less than 14 days in 48 cases(64.9%),more than 14 days in 26 cases(35.1%).26 cases(35.1%) were diagnosed by sputum smear examination.27 cases(36.7%) were diagnosed by biopsy methods(8 cases by biopsy via bronchoscope,17cases by open-thorax lung biopsy, 1 case by lung-puncture biopsy and 1 case by lymphonode biopsy).The positive rate was 100% for every biopsy method except bronchoscopic biopsy(88.9%).20 cases(27.0%) were clinically diagnosed. Conclusion Respiratory department of general hospital is on the first-line of pulmonary tuberculosis prevention.The detection level of pulmonary tuberculosis in people’s Hospital of Peking University is satisfactory,and most patients were diagnosed in two weeks.The main diagnostic method is sputum smear examination,but a few parts of cases were diagnosed through biopsy methods.

Evaluation of the clinical safety of Yifeizhike capsule

Study group for clinical safety of Yifeizhike capsule

. 2007, 29(5):  415-418. 

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Objective To evaluate the clinical safety of Yifeizhike capsule in treatment of tuberculosis. Methods Two hundred and fifty new patients with pulmonary tuberculosis were randomly divided into observation group(125 cases) and control group(125 cases).The observation group received chemical therapy and Yifeizhike capsule, while the control group only received the same chemical therapy.Evaluate the adverse drug reaction and the abnormity of blood,urine and the function of liver and kidney within 8 weeks. Results There was no significant difference between the two groups in white blood cell(WBC),red blood cell(RBC),platelet(PLT),glutamic pyruvic transaminase(ALT),glutamic oxalacetic transaminase(AST),blood urine nitrogen(BUN) and uric acid(UA);and there was no significant difference between the two groups in digestive system,allergic reaction,nervous system and arthralgia either.6 cases stopped using drugs because of the adverse drug reaction in each group respectively,11 cases were due to using the anti-TB drugs and 1 case was due to using Yifeizhike capsule with the occuring of allergic skin. Conclusion Yifeizhike capsule is safe in clinical use,and the problem of allergic skin should be paid more attention.

Clinical observation on the clinical efficacy of compound Glycyrrhizin in the treatment of new pulmonary tuberculosis complicated with chronic hepatitis B

Weng Lizhen,Zhu Yufang,Chen Shuqin,et al.

. 2007, 29(5):  419-422. 

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Objective To observe the clinical efficacy and the adverse reactions of Compound Glycyrrhizin(Stronger Neo-Minophagen C) in the treatment of new pulmonary tuberculosis complicated with chronic Hepatitis B. Methods Sixty cases of new pulmonary tuberculosis complicated with chronic Hepatitis B were randomly divided into the treatment group(30 cases) and control group(30 cases).The both groups were treated with effective antitubercuolsis drugs.The treatment group was treated with Stronger Neo-Minophagen C,while the control group took glutathione. Results After two months treatment,the total effective rate of liver function improvement was 76.7% in the treatment group,and 46.7%(P<0.05) in the control group.The rates of clinical symptom and physical sign disappearance as well as distinct amelioration were 40% and 56.7% in the treatment group,and 26.7% and 46.7% in the control group,respectively.There were notable differences between the treatment group and the control group in the above mentioned aspects.Adverse reactions were rarely found. Conclusion Stronger Neo-Minophagen C is effective and reliable in improving liver function of patients suffering from new pulmonary tuberculosis complicated with chronic Hepatitis B during anti-tuberculosis chemotherapy.

Study on the activity of the Fixed-dose combination in antituberculous

Li Huiwen,Liang Guifang,Zhao Weijie,et al.

. 2007, 29(5):  423-428. 

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Objective To compare the antituberculous activities of the Fixed-dose combination(FDC) in antituberculous made by different pharmaceutical companies and evaluate their potential contribution to treatment of tuberculosis. Methods The MICs of the FDC against M.tuberculosis H₃₇Rv、M.phlei and M.ravenal were determined by the tube doubling dilution method in vitro.The efficacy of antituberculosis was assessed by half survival time of the mice. Results In vitro, the MICs of main composition in the FDC(RFP,INH or RFP,INH PZA) against M.tuberculosis H37Rv、M.phlei and M.ravenal were lower than that of each composition used alone in the same media.In vivo,these FDC drugs had marked effect against experimental tuberculosis in mice and were superior to each agent used alone with the same dose. Conclusion The two-and three-drug FDC had significantly effective antituberculous activity.The same preparations from different companies had not significant difference in treatment of the mice infected with M.tuberculosis.

The CT features of tuberculosis in hip joint

He Wei,Xu Jingping,Zhou Xinhua.

. 2007, 29(5):  429-431. 

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Objective To evaluate the CT features of tuberculosis in the hip joint. Methods The 41 cases who were diagnosed definitely as tuberculosis in the hip joint were analyzed the CT appearances,and compared the CT appearances in children with that in adults. Results The CT main features of tuberculosis in hip joint were as follow: bone destruction(90.3%),hip and lower limb’s muscles atrophy(87.7%),soft tissue swelling and abscess(75.6%),joint space widened or narrowed(73.1%),joint space effusion(63.5%) and juxta-articular osteoporosis(46.3%).Joint space effusion in children was more than that in adults. Conclusion CT examination had an important value in the diagnosis of tuberculosis in hip joint.

Clinical analysis of 5 cases of pulmonary vasculitis misdiagnosed as pulmonary tuberculosis and literature review

Liu Qiangui¹,Sun Li,Zhao Shuangyan,et al.

. 2007, 29(5):  432-434. 

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Objective To discuss the causes and prevention measures of misdiagnoses of pulmonary vasculitis as pulmonary tuberculosis. Methods Through discussion of the causes of misdiagnoses of 5 cases pulmonary vasculitis as pulmonary tuberculosis,we put forward the correct thinking way of diagnosis and misdiagnosis prevention Methods for pulmonary vasculitis. Results Five patients with pulmonary vasculitis all were misdiagnosed as pulmonary tuberculosis,and received non-proper examination and therapy,delayed disease diagnosis and treatment.3 patients were improving and discharged,and 2 patients died. Conclusion clinical physicians should raise their understanding of pulmonary vasculitis.When the patients don’t show improvement after relevant treatment,the anti-neutrophil cytoplasmic autoantibody(ANCA) in serum should be examined in time.If there are good conditions,lung biopsy should be done to diagnose definitely.