Table of Content

10 May 2012, Volume 34 Issue 5

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Review of the number and survey method of people infected with Mycobacterium tuberculosis in China

XU Miao, WANG Guo-zhi

Chinese Journal of Antituberculosis. 2012, 34(5):  269-270. 

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Study on the reliability of detecting Mycobacteria in sputum specimens by PCR-fluorescent probe

LIANG Jian-qin， GAO Hua-fang， LI Hong-min， ZHAO Zhi-xian， ZHANG Guang-yu， WANG Jin-he， WANG Su-min， LIU Jia-wen

Chinese Journal of Antituberculosis. 2012, 34(5):  271-274. 

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Objective  To evaluate the clinical application value of detecting the Mycobacterium tuberculosis and non-tuberculous mycobacteria (NTM) by PCR-fluorescent probe.  Methods  1015 sputum specimens and 56 clinical isolates were studied using PCR-fluorescent probe of Capital Bio Corporation. And the results were compared with that of Mycobacterium tuberculosis detection kit of Daan Company.  Results  The sensitivity and specificity of detecting Mycobacteria in the sputum specimens by PCR-fluorescent probe were 50.3% (373/741) and 98.9% (271/274) respectively, in which the sensitivities for smear-positive and smear-negative pulmonary tuberculosis were 97.0%（257/265） and 24.4%（116/476） respectively. Of 1015 sputum specimens, 11（1.1%）were detected as NTM, Using 16S rDNA sequencing as control, the accuracy rate was 100.0%. 138 sputum specimens were detected repeatedly, the coincidence rate was 100.0%. Of 56 clinical isolates, 1 strain was detected as Mycobacterium tuberculosis and 55 were detected as NTM, the accuracy rate was also 100.0%. The total coincidence rate of 226 sputum specimens was 95.1% (215/226) compared with Mycobacterium tuberculosis detection kit of Daan Company. There were no significant difference between two methods (χ²=2.98, P=0.226). Conclusion  PCR-fluorescent probe can rapidly detect and differentiate Mycobacterium tuberculosis and NTM, have the value of clinical application.

Study on the molecular mechanism of mono-rifampicin resistance in Mycobacterium tuberculosis

PANG Yu, LI Gui-lian, WANG Yu-feng, SONG Yuan-yuan, LI Qiang, OU Xi-chao, DONG Hai-yan, ZHAO Yan-lin

Chinese Journal of Antituberculosis. 2012, 34(5):  275-279. 

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Objective  To investigate the roles of drug-resistant related gene mutation and efflux pump genes in mono-rifampicin resistance of Mycobacterium tuberculosis isolates.  Methods Twenty-three mono-rifampicin resistant clinical isolates identified in national survey of drug-resistant tuberculosis in 2007—2008 were obtained from National Tuberculosis Reference Laboratory. Direct sequencing was performed to detect the mutations in the rpoB gene. The total RNA of the rifampicin resistant strains without rpoB mutation was extracted and reverse-transcribed. The transcript level of 20 drug efflux genes was detected by real-time PCR and the RFP-related efflux pumps were screened by comparing with the corresponding results of minimum inhibitory concentration (MIC). The function of candidates in RFP-resistance was confirmed by transgenic E.coli.  Results Of 23 mono-rifampicin resistant strains, the mutations of rpoB were detected in 16 (69.6%) strains, including 9 isolates with mutation located in 531 codon, 6 isolates in 526 codon and 1 isolate in 533 codon. The isolates with Ser531Leu and His526Asp mutation displayed high-level rifampicin resistance, the MIC of which was 192 μg/ml and 256 μg/ml, respectively, while the mutations including His526Gly，His526Leu，His526Arg and Leu533Pro were related to low-level rifampicin resistance, the MIC of which was 0.75 μg/ml，4 μg/ml，2 μg/ml and 0.5 μg/ml, respectively. In the strains without rpoB mutation, the expression levels of Rv2936, Rv0783 and Rv0933 by PCR positively correlated to MIC. Moreover, the BL21 transferred with pEASY-E1-Rv2936 and pEASY-E1-Rv0783 showed higher MIC level, 32 μg/ml and 16 μg/ml, respectively, than 8 μg/ml in the control E.coli and the E.coli transferred with pEASY-E1-Rv0933.  Conclusion The different mutations in rpoB are associated with MIC level. And Rv2936 and Rv0783 may be the efflux pumps related to rifampicin resistance in Mycobacterium tuberculosis.

The value of FQ-PCR for efficacy evaluation in children tuberculous meningitis

LIU Yan-xuan, JIA An-kui, GUO Sheng-ju, SHI Tai-xin, SHANG Hao-zhen

Chinese Journal of Antituberculosis. 2012, 34(5):  280-283. 

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Objective  To explore the roles of FQ-PCR in rapid diagnosis，efficacy evaluation and prognosis for children tuberculous meningitis（TBM）. Methods  A total of 147 patients of children were enrolled in the study, 117 of whom were confirmed in prodromal stage TBM by the gold standard(culture with MGIT ^TM 960) and the others(n=30) were control group without TBM. Cerebrospinal fluid（CSF）specimens of 147 cases were immediately numbered and detected by FQ-PCR and then the sensitivity(SE), specificity(SP), diagnostic index(DI), positive predictive value(PPV), negative predictive value(NPV), agreement rate(AR) and other statistical indicators were calculate and analyzed. CSF of TBM group were cultured blindly and detected by FQ-PCR after 3 weeks and the two detection rate were calculated. Patients discharged from hospital who presented with negative by the two methods in the TBM group were followed up for 2 years and the recurrence rate was calculated.  Results  Compared to the gold standard, the SE，SP，DI and AR of FQ-PCR was 100.0%(117/117), 96.7%(29/30), 196.7%(116/117), and 99.3%, respectively(χ²=0.000, P=1.000). There was a correlation between the two methods(r=0.9790, χ²=140.9, P=0.000). Thirty-two cases and 68 cases of the TBM group were detected by culture method and FQ-PCR after treatment for 3 weeks, respectively(χ²=22.63, P=0.000). The follow-up results showed that 7 cases of 78 patients with negative culture result relapsed, 69 patients with negative FQ-PCR had no recurrence,（χ²=4.6736, P=0.0306）.  Conclusion  FQ-PCR detection of cerebrospinal fluid TB-DNA is accurate, simple, rapid method, which can be used for early diagnosis of tuberculous meningitis, but also has advantages of prognosis monitoring and efficacy evaluation than the culture method.

Clinical analysis of 643 elderly pulmonary tuberculosis

SU Rui，WANG Zhong-yuan，CHEN Hong-bing，LIU Lin，ZHANG Tao，AN Hui-ru，WANG Tao

Chinese Journal of Antituberculosis. 2012, 34(5):  284-288. 

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Objective  We analyzed the clinical characteristics of elderly pulmonary tuberculosis (TB) in order to improve the diagnosis and treatment of this disease.  Methods  We retrospectively analyzed their clinical presentation, radiological features，sputum （or BALF） AFB，underlying diseases，anti-TB adverse effects and the responses to anti-TB chemotherapy among 643 hospitalized elderly cases with pulmonary tuberculosis in the 309th hospital.  Results  This group had 93.9% (604/643) secondary tuberculosis,6.1% (39/643) hematogenous dissemination,and no primary tuberculosis. The rate of the positive AFB including smear and culture from both sputum and BALF together was 31.9% (205/643). Coughing was the most common symptom (79.9%,514/643) which appeared more often in the positive AFB cases than that in the negatives（87.8% vs. 76.3%; χ²=11.614，P=0.001）. On the other hand, night-sweat appeared more common in the negatives (32.9% vs. 24.4%; χ²=4.774, P=0.029). Almost 90% of them had underlying diseases, 78.1% (450/576) had at least two kinds. The most common disease was pneumonia (51.2%,329/643) which was more popular in the positive AFB cases than in the negatives (57.1% vs. 48.4%; χ²=4.202, P=0.040).The hypertension was more common in females than in males (19.7% vs. 13.2%; χ²=4.380, P=0.036).Infiltration and necrosis were more common imaging in 604 case with secondary tuberculosis and multiple lung  fields were involved (41.4％，250/604). 59.5% (122/205) of them became sputum conversion negative after treatment during the hospitalization. The effective rate of this short-term chemotherapy was 87.1% (560/643), and the mortality rate was 5.1% (33/643). The most prevalent adverse effects of anti-TB chemotherapy were liver toxicity (6.4%,41/643).  Conclusion  The elderly pulmonary tuberculosis presented more with coughing, fever,some underlying diseases including the pneumonia as the top one.Their pulmonary focuses were characteristic with distributing in one more lung fields. Elderly pulmonary tuberculosis should be given personalized treatment.

Cross-sectional survey on pulmonary tuberculosis in two prisons of Hubei

XIONG Chang-fu, ZHOU Li-ping, WANG Ming-qiu, LI Ai-guo, SHA Sha, LIU Xun, WANG Xiao-jing, DING Xue, LIU Xiao-jun

Chinese Journal of Antituberculosis. 2012, 34(5):  289-292. 

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Objective  To understand the situation of TB epidemic in prison and the status of prevention work.  Methods The pulmonary tuberculosis (PTB) census and questionnaire investigation were performed for all the prisoners, and sputum smear, sputum culture, bacterial identification and drug susceptibility test (DST) were done for prisoners with abnormal chest radiograph shadow or suspicious symptoms. Besides, the infectious source homology analysis was made for culture positive patients through Mycobacterial interspersed repetitive unit-variable number of tandem repeat (MIRU-VNTR) typing method. The prevalence of tuberculosis in prison was calculated and compared with the general population.  Results  A total of 40 cases were diagnosed as PTB, of which one was sputum smear positive, one was culture positive (same with the smear positive), five were smear negative and culture positive, and 34 were smear negative and culture negative. The DST results for six culture positive patients showed that, there are three drug-resistant cases, among which, one was streptomycin-resistant, one was resistant to isoniazid, rifampicin and ethambutol simultaneously, one resisted to isoniazid, rifampicin, ethambutol and ofloxacin. The prevalence of PTB, smear positive, culture positive were 1156.4/100 000（40/3459）, 28.9/100.000（1/3459）, 173.5/100 000（6/3459） respectively. The PTB prevalence in prisoners was higher than that in local residents(χ²=17.858, P<0.01).The total awareness rate for three pieces of core TB information was 61.6%（6384/10 359）， while 67.4%（4737/7026） for the B prison，49.4% （1647/3333）for the A prison. The difference was statistically significant (χ²=309.1，P<0.01). In addition, the homology analysis showed that the infectious sources of the five culture-positive patients in the same prison were different.  Conclusion  The TB epidemic in prison is serious, so the effective prison TB control working model should be explored.

Analysis of needs and utilization of health services in patients with pulmonary tuberculosis in Minhang district, Shanghai

XIA Tian, TANG Li-hong, CHENG Yu-ping, SHEN Xin, MEI Jian, SHEN Mei, YUAN Zheng-an, SONG Yuan-cheng, XING Bo

Chinese Journal of Antituberculosis. 2012, 34(5):  293-298. 

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Objective  To analyze the needs and utilization of health service in patients with pulmonary tuberculosis in Minhang district of Shanghai city, and evaluate the factors influencing the utilization of health service.  Methods  In 2010, a total of 1 286 830 copies of electronic health records were established in Minhang district, of which 749 were records specially for registered pulmonary tuberculosis. These 749 pulmonary tuberculosis cases and 502 patients among them which have been cured were taken as research subjects to analyze their epidemiological and management information, the needs and utilization of health service for tuberculosis patients. Univariate and multivariate logistic regression analysis was made to analyze the factors affecting their health service utilization. Results  Among the filing population in Minhang district in 2010, the prevalence of pulmonary tuberculosis was 58.21/100 000, the prevalence of active pulmonary TB (sputum positive) was 30.62/100 000, and the prevalence of chronic diseases was 51.54%（386/749）. Among the 502 cases who have been cured or have completed the full-course treatment, the frequency of clinic visits, laboratory examinations, and DOT supervisions in terms of times were 2.74, 9.02 and 5.47 times respectively. Patients older than 65 years (β=－2.513, Wald χ²=0.508, P=0.000, OR(95.0%CI)=0.081(0.030~0.219)), with pulmonary tuberculosis and other chronic diseases(β=－3.987, Wald χ²=0.458, P=0.000, OR(95.0% CI)=0.136(0.047~0.491)) are more likely to increase the frequency of clinic visits. Conclusion  The patients with pulmonary tuberculosis in Minhang district have a high requirement for health services, however, the utilization is adversely low. We should carry out targeted prevention measures for different groups of people, grasp their needs of health service, and improve their health services utilization.

Analysis of the case detection and short-term treatment effect of the Wuhan MDR-TB project

ZHOU Mei-lan, CHEN Zi, WANG Jian-jie, CHEN Cong, PAN Hong, WANG Wei-hua

Chinese Journal of Antituberculosis. 2012, 34(5):  299-303. 

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Objective  To analyze and evaluate the case detection and short-term treatment effect of the Wuhan MDR-TB project, and provide reference for MDR-TB control.
Methods  The baseline information, data reports and case medical records between December 2006 and June 2010 was collected for analysis and evaluation. A total of 998 smear positive patients dropping into four categories of registration-chronic, initial treatment failure, relapse and other relapse were screened and  893 cases have been identified as MTB infection.  Results  Among the 893 cases, 127 cases were chronic patients, 59 cases were initial treatment failure patients, 451 cases were relapse patients and 256 cases were other relapses. All of them were administered to drug susceptible test (DST), 229 cases (25.6%, 229/893) were confirmed as MDR-TB cases; the MDR-TB detection rate for chronic patients, initial treatment failure patients, relapse and other relapse patients were 35.4% (45/127), 33.9% (20/59), 26.4% (119/451) and 17.6% (45/256) respectively; In terms of MDR detection rate, there is no significant difference between chronic and initial treatment failure patients (χ²=0.04, P＞0.05)，however, these rate are notably higher than relapse and other relapse patients(χ²=3.99, χ²=15.05, P＜0.05); among 229 cases of confirmed MDR-TB, 128 cases(55.9%，128/229) had been treated with MDR-TB regimens; the adverse reaction incidence was 85.2% (109/128); sputum smear and culture conversion rates at the end of 6-months treatment was 87.5% (98/112), 89.3% (100/112) respectively; 43 cases of MDR-TB had completed the full course of treatment, the cure rate was 60.5% (26/43).  Conclusion  Chronic, initial treatment failure and relapsed smear positive patients are main source of MDR-TB patients. MDR-TB patients have a low treatment reception rate and cure rate, high incidence of adverse reaction. Prevention and treatment are equally important in MDR-TB control.

Controlled clinical studies comparing the new chemotherapeutic regimens with the primary regimens for retreatment pulmonary tuberculosis patients

ZHU Li-zhen,GAO Meng-qiu,CHEN Wei,LI Fang,LI Zhi-hui,YE Zhi-zhong,SHI Jun-wei,LI Li,YUE Ji,SHEN Yun-fei,ZHANG Li-qun,YANG Guo-feng,LI Guang-zhong,GAO Yuan,QIU Li-hua,ZHANG Hong-man,WANG Sheng-wei,WU X

Chinese Journal of Antituberculosis. 2012, 34(5):  304-309. 

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Objective  To evaluate the efficacy and safety of new chemotherapeutic regimens for retreatment pulmonary tuberculosis. Methods  A total of 345 cases were enrolled，according to the results of drug susceptibility testing, 161 cases with drug-sensitive results were assigned to a test group（85 cases） and a control group (76 cases),184 with drug-resistant results assigned to the test group（124 cases） and the control group (60 cases). The chemotherapeutic regimens were as follows：(1)For the drug-sensitive group, new chemotherapeutic regimen with 2HRZES/6-10HRE was used in the test group and primary regimen with 2H₃R₃Z₃E₃S₃/6H₃R₃E₃  was used in the control, respectively(8 months for PTB but no diabete cases,12 months for PTB with diabete cases). (2)For the drug-resistant group, 3R（H）ZES±Lfx/6-9R(H)ZE±Lfx was used in the test group. R or H could be replaced each other for R or H resistant patients. Am could be used in the case of S resistance. Am and Lfx could be used in HS or RS resistant cases. 2-3HRZES/6HRE regimen was used in the control(We selected the control subjects in the same place, same situation but historical controls). Results  (1)In drug sensitive group, the sputum conversion rate was 77.6%(66/85) and 88.2%（67/76）, radiographic improvement rate was 63.5%(54/85) and 55.3%（42/76）, effective rate was 96.5%(82/85) and 90.8%（69/76）and cavity closing rate was 68.3%(28/41) and 56.1%（23/41）,there was no statistically significant difference between the test group and the control group（χ²＝2.397 24，χ²＝0.821 42，χ²＝1.354 777，χ²＝0.829 58, P＞0.05）. Also in drug sensitive group complicated with diabetic, the sputum conversion rate was 66.7%（16/24） and 73.3%（11/15）, radiographic improvement rate was 58.3%（14/24） and 46.7%（7/15）,there was no statistically significant difference between the test group and control group complicated with diabetic（χ²＝0.191，χ²＝0.511，P＞0.05）. Meanwhile, the sputum conversion rate was 82.0%（50/61） and 91.8%（56/61）, radiographic improvement rate was 65.6%（40/61） and 57.4%（35/61）in the test group but no diabetes and control group but no diabetes. There was no statistically significant difference between the two groups（χ²＝2.589 62，χ²＝0.861，P＞0.05）.（2）In the drug resistant group,  at the end of treatment, the radiographic improvement rate was 79.8%（99/124）in the test group and 66.7%(40/60) in control. There were statistically significant difference between the test group and control （χ²＝3.81, P＜0.05）;Radiographic improvement rate was 62.9%(78/124) in test group and 45.0%(27/60) in the control, there were statistically significant difference between the test group  and the control（χ²＝4.584 29, P＜0.05）; The Incidence of adverse reactions were 1.1%（1/92） and 1.2%(1/82) in the test and control groups in drug sensitive group, respectively, and the Incidence of adverse reaction in the test group of drug-resistant cases was 2.8％（4/142）. Conclusion  For retreatment drug-sensitive cases, whether or not complicated with diabetes, can receive the primary chemotherapeutic regimen. New chemotherapeutic regimens had got good effect in the retreatment drug-resistant cases and the Incidence of adverse reactions were lower and no sever adverse reactions developed.

Analysis on the implementation status of cross-regional management procedures of pulmonary tuberculosis patients in 4 provinces

LI Jun，JIANG Shi-wen，HUANG Fei，LI Xue，ZHANG Hui，LIU Xiao-qiu

Chinese Journal of Antituberculosis. 2012, 34(5):  310-314. 

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Objective  To investigate and evaluate the implementation status of cross-regional management procedures of pulmonary tuberculosis patients, and to provide scientific evidence to further improve the nation-wide cross-regional management procedures and work quality.  Methods  In terms of the number of patients transferred in and out between July to December in 2009, the top 4 provinces namely Guangxi, Guangdong, Fujian and Zhejiang province were selected for field study. In turn, each province selected top counties as study site. A combination of quantitative and qualitative method was adopted to conduct surveys and on-site interviews within the selected counties. There were totally 16 counties for field study and 40 respondents involved for interview.  Results  As on-site survey from the place the patients transferred out indicated, there were still 9.4%(45/479) of patients taking their drug and sputum examination in their first-registered dispensaries. Patients transferred through TB information management system accounted for 79.3 %(344/434) of actual patients transferred. The consistency rate was 84.8%(875/1032） between actual patient roll-out information and information from TB management system. As on-site survey from the place that patients transferred to illustrated, patients’ feedback information was highly consistent  with reports from TB information management system; while sputum examination results and treatment outcome feedback showed low consistency, which were 61.5%(198/322) and 45.6%(52/114) respectively. Some of the information had not been fed back by TB information management system in a timely manner. Interview results showed that the nation-wide cross-regional management procedure of pulmonary tuberculosis patients and TB management system had been generally recognized by respondents in the study areas, and could meet the management requirement. However there were still some difficulties and problems during the implementation process.  Conclusion  According to the key points of procedure and identified problems, more specific and focused working measures are demanded which turns out to further improve the performance quality of cross-regional pulmonary tuberculosis patients’ management.

Role of real-time fluorescence PCR in Mycobacterium tuberculosis complex DNA detection from stool samples of children with pulmonary tuberculosis

HUANG Hui-qian, WU Chi, CHEN Jian-bo, YANG Yan, HOU Zhi-ping, LI Xiao-yong

Chinese Journal of Antituberculosis. 2012, 34(5):  315-318. 

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Objective  To establish a rapid detection process of M. tuberculosis DNA in stool specimens of children using real-time fluorescence PCR and investigate its clinical diagnostic value.  Methods  We collected 76 stool specimens of hospitalized children with pulmonary tuberculosis and detected the M. tuberculosis DNA using real-time PCR to compare the results with the PCR results of 20 stool samples of children with other pulmonary diseases, 76 stool smear results and 41 sputum samples’ smear and (or) culture, and real-time PCR results.  Results  The sensitivity of real-time PCR in stool samples was 23.68%(18/76), and the specificity is 100.00%（20/20）. The positive rate of stool PCR (23.68%,18/76) was significantly higher than the smear test (6.58%,5/76). The number of cases tested positive from sputum smear and (or) culture (15 cases) and sputum PCR (18 cases) were larger than the number of cases tested positive from fecal PCR (11 cases).  Conclusion  Using real-time PCR to find TB-DNA in stool samples of children, is a highly specific, moderately sensitive, non-invasive and relatively safe method for the diagnosis of children pulmonary tuberculosis.

Analysis of awareness rate of tuberculosis prevention knowledge among the floating population in Shandong province

WEI Qian, WANG Shi-chang, LOU Lei, WANG Zhong-dong, MA Yun-bo, LIU Gui-qiang, LIU Cun-tang

Chinese Journal of Antituberculosis. 2012, 34(5):  319-323. 

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Objective  To understand the status of awareness of TB knowledge and its influence factors, and to provide a reference for floating population health education and health promotion.   Methods  Four hundred and forty-seven floating people were selected through the cluster random sampling method, and they were administered to face-to-face questionnaire survey. Four hundred valid questionnaires were collected eventually, and the effective rate is 89.5%(400/447). The Epidata 3.1 software was used to set up the database and double entry of the data, Excel 2003 and SPSS 13.0 were used for data analysis.   Results  The overall awareness rate to core information in all respondents is 57.1 % (1141/2000), 76.9 %(684/890) in male and 41.2 %(457/1110) in female respectively. This rate is significantly higher in male than in female（χ²=256.67，P＜0.01）. In terms of age-specific TB awareness rate, highest rate is 69.8 %(199/285) in 30- age group, followed by 64.0%(285/445) in 40-50 age group. The lowest rate is 51.7%(657/1270) in 18- age group. The difference is statistically significant（χ²=42.60，P＜0.01）. Regarding the proportion of respondents by age, the highest proportion is 18- age group, accounting for 63.5%(254/400)of all the respondents. The awareness rate is 59.4 %(692/1165) in migrant workers who have been worked for ≥2 years in surveyed locations, higher than those worked for <2 years which is 53.8%(449/835)（χ²=6.28，P<0.05）. In 5 core information, one specified as “cough, expectoration more than 2 weeks, or blood stained sputum, should be suspected of suffering from TB” has the lowest awareness rate, 36.0%(144/400) only. 67.5%(239/354) of respondents acquired TB knowledge by TV, 46.6%(165/354) were told by others.   Conclusion  The awareness rate of TB knowledge in the floating people is still low in Shandong province. We should strengthen TB health education for new generation of floating population and new migrant workers, especially focusing on the dissemination of important core information.

Research progress of new drugs for anti multi-drug resistance tuberculosis (MDR-TB)

ZHAO Heng-min, WANG Wei-hua

Chinese Journal of Antituberculosis. 2012, 34(5):  324-326. 

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