Table of Content

10 August 2012, Volume 34 Issue 8

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The fifth national tuberculosis epidemiological survey in 2010

Technical Guidance Group of the Fifth National TB Epidemiological Survey；The Office of the Fifth National TB Epidemiological Survey

Chinese Journal of Antituberculosis. 2012, 34(8):  485-508. 

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This survey aims to understand the prevalence status and trend of tuberculosis (TB) in China, and evaluate the implementation of National TB Control Programme (2001—2010). Multi-stage stratified cluster proportional random sampling method was used to select the survey population. People over 15 years old in sampled survey points were administered chest X-ray, those with suspected pulmonary TB symptoms or with suspected pulmonary TB lesion shown by X-ray were performed sputum smear and culture examination. All the bacterial strain obtained were performed drug susceptibility testing of anti-TB drugs. All active TB patients detected by this survey received the socio-economic and TB knowledge awareness questionnaire. This survey has following findings. First, the prevalence of active and smear positive was 459/100 000 and 66/100 000 respectively among population over 15 years old in 2010. Secondly, the prevalence in male was higher than in female, and gradually increase by age, peaked in 75-79 years old. Thirdly, the active and smear positive prevalence of pulmonary TB were 291/100 000 and 44/100 000 in eastern part of China, 463/100 000 and 60/100 000 in the middle part, 695/100 000 and 105/100 000 in western part, 569/100 000 and 78/100 000 in rural area, 307/100 000 and 49/100 000 in cities respectively. Fourth, the multi-drug resistance TB rate was 6.8% (19/280). Fifth, the general pulblic TB knowledge awareness rate was 57.0% (720 912/1 264 905). The sixth, the annual per capital net income of TB patient household was 3292 yuan, of those 66.8% of patients lower than 60% of nationwide per capita income. In compariton with the survey in 2000, the prevalence of smear positive pulmonary TB showed a downward trend among people over 15 years old, as well as in different age group and gender. However, the prevalence in rural area was higher than in cities, and western part also significantly higher than the middle and eastern parts. These findings indicated that although the TB prevalence has dropped, the TB burden especially for the multi-drug resistant TB is still very serious.

Health care seeking behavior among pulmonary tuberculosis suspects in Wuhan city: a community-based study

DUAN Qiong-hong, WANG Peng, Lv Jing, ZHONG Rong, WANG Wei-hua, ZHOU Zhi-hua, ZENG Qing-zhi

Chinese Journal of Antituberculosis. 2012, 34(8):  509-513. 

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Objective  To investigate the patterns of health care seeking behavior and to identify the factors affecting the behavior among community TB suspects in Wuhan city, and to provide scientific basis for improvement of TB case detection.   Methods  We followed a cluster proportional random sampling method to select the 4 study communities (clusters), a total of 5878 people above 15 years old. A total of 270 people who had continuous cough, expectoration for more than two weeks, hemoptysis or who had bloody sputum within six months prior to the survey enrolled in the survey. All the participants received questionnaire interview and chest X-ray, sputum smear and culture examination. Two hundred and fifty-nine pieces of questionnaires were qualified with the 95.9% percent of pass. The chi-square test and logistic regression were used to analyze the factors affecting the health care seeking behavior among TB suspects.  Results  Among the 259 participants with eligible questionnaires, 79 cases (30.5%) did not seek any health care, 86 cases (33.2%) sought medical services from primary health care centers or general hospitals，89 cases（34.4%）had self-medication，while 5 cases（1.9%）had gone to private clinics. Multivariate logistic regression analysis showed that the availability of health insurance（β=1.225，Wald χ²=3.955, OR=3.405, 95%CI＝1.018-11.392, P<0.05）, the presence of severe or chronic respiratory symptom（β=1.601，Wald χ²=22.210，OR=4.959, 95% CI＝2.548-9.652, P<0.01）, and the feeling of stigmatization (β=0.927，Wald χ²=7.767，OR=2.528, 95%CI＝1.317-4.853，P<0.01) were correlated to the health seeking behavior to above community level health service center.  Conclusions  The proportion of TB suspects who actively sought health care was low. The health insurance, severe or chronic respiratory symptoms and stigmatization were the main factors affecting the health care seeking behavior among community TB suspects.

An analysis of 321 cases of multidrug-resistance tuberculosis patients

HE Pei-xian, LI Wan-mei, KANG Li-juan

Chinese Journal of Antituberculosis. 2012, 34(8):  514-518. 

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Objective  To understand the age distribution of primary and acquired MDR-TB patients and the status of their drug resistance, and provide a scientific basis for prevention and treatment of MDR-TB.  Methods  We made a retrospective analysis of MDR-TB cases admitted in our hospital between January 2005 and December 2010 (47 cases of primary and 274 cases of acquired  multidrug-resistant cases respectively). Patients were divided into 4 groups by age, 5 cases in ＜20 age group, 183 cases in 20- age group, 105 cases of  40- age group, and 28 cases of 60- age group, respectively, to analyze the status of TB drug resistance.  Results  In 47 primary and 274 acquired MDR cases,age between 20 to 30 had the highest number of cases, which were 30(63.8%)and 153 (55.8%) respectively. However the difference of primary and acquire MDR-TB distribution in different age group is not statistically significant(primary MDR-TB: 1 case of ＜20 age group (2.1%), 14 cases of age over 40 (29.8%), 2 cases of age over 60(4.3%), acquired MDR-TB: 4 cases of ＜20 age group(1.5%), 91 cases of age over 40 (33.2%),  26 cases of age over 60 (9.5%). (Fisher’s exact test, P=0.515). As for the distribution of the first-line drug resistance (respectively, resistance to the HR, HRE, HRS and HRSE), in primary MDR cases, 1 cases in ＜20 age group (resistant to HRS), 30 cases of age over 20 (10 resistant to HR, 13 to HRE，5 to HRS, and 2 to HRSE), 14 cases of age over 40 (4 resistant to HR, 7 to HRE, 3 to HRS), 2 cases of age over 60 (1 resistant to HRE and 1 to HRS). In acquired MDR-TB cases, 4 cases in ＜20 age group (resistant to HR, HRE, HRS and HRSE respectively), 153 cases of age over 20 (56 (36.6%) resistant to HR, 26(17.0%) to HRE，46(30.1%) to HRS, and 25(16.3%) to HRSE), 91 cases of age over 40 (34(37.4%) resistant to HR, 26(28.6%) to HRE, 17(18.7%) to HRS, 14(15.4%)to HRSE), 26 cases of age over 60 (5(19.2%) resistant to HR and 5(19.2%)  to HRS, 13(50.0%)to HRE, 3(11.5%) to HRSE). The distribution is statistically significant different between different age group (Fisher’s exact test, P=0.030).Further multiple comparison found that the difference is statistically significant between 20- age group and 60- age group. In primary MDR and acquired MDR cases, they were mainly resisted to one or two second-line drugs(seven drugs in total: amikacin (Am), clarithromycin (Clr), levofloxacin (Lfx), moxifloxacin (Mfx), sodium aminosalicylate (PAS), rifabutin (Rfb), propylthiouracil isonicotinoyl amine (Pto)］, the resistant rate were 25(53.2%) and 115(42.0%) respectively, however the difference is not statistically significant (χ²=3.123, P=0.211).  Conclusion  The primary and acquired MDR-TB predominantly affects young people, and the situation of MDR-TB is serious. We should strengthen the diagnosis and treatment of TB patients, and reduce the incidence of multi-drug-resistant.

The analysis of self-efficacy and its influence factors in pulmonary tuberculosis cases

LI Gui-qing, TAN Shou-yong, GAO Cui-nan, HE Qing-qiu, DENG Hong

Chinese Journal of Antituberculosis. 2012, 34(8):  519-522. 

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Objective  To evaluate the status of self-efficacy and its influence factors in pulmonary TB cases, then explore an effective approach to improve their self-efficacy levels.  Methods  A total of 182 patients with pulmonary TB were sampled by random number table method. General social-demographic questionnaire and self-efficacy questionnaire were used to investigate patients’ general information and self-efficacy in this study. One hundred and eighty-two pieces of questionnaires were issued and returned with the valid rate of 100.0%. Self-efficacy of pulmonary TB is divided into high, medium and low three levels. ≥ 34 is considered high level, and 25～33 as medium level, ≤ 24 as low level.  Results  The total score of self-efficacy in 182 patients with pulmonary TB was (26.45±4.99). 64.8% (118/182) of patients had a medium level of self-efficacy, Only 17.6%(32/182) of patients were in high level. Male scored (24.35±4.84), the female scored (26.74±5.19) (t=37.654, P<0.01); Those who had a personal monthly income of less than 2000 Yuan scored  (27.15±3.56), those between 2000 to 5000 scored （27.46±5.46）, and ＞5000 scored （25.32±4.54）(F=3.884, P<0.05); Patients who had a good family and social support scored (27.14±4.92), and （25.68±5.01）for those with low support（t=12.728，P＜0.01）. Patients with good psychological condition scored (26.92±5.00) and （26.53±5.07）for bad condition（t=6.284，P＜0.01). In terms of education level, patients with college level scored （27.48±4.68）, high school scored (26.32±4.91), junior high school scored (27.64±5.53) and primary school score (24.70±4.24) (F=2.837, P<0.05); Patients with positive attitude towards disease scored （26.86±4.95）, ‘does no matter’attitude scored （24.53±4.49）（t=31.042，P＜0.01）.  Conclusion  The self-efficacy in patient with pulmonary TB was affected by many factors. Therefore, we should focus on main influencing factors to take appropriate measures in order to improve self-efficacy of pulmonary TB.

Cloning, expression, purification and identification of Rv3803c, Rv3846 gene from Mycobacterium tuberculosis

SUN Yi-fan，LI Hai-cheng，ZHOU Jie，QIAN Ming，CHEN Tao，JIANG Yong，LAI Sai-lin，JIANG Zhen-you，ZHONG Qiu，ZHOU lin

Chinese Journal of Antituberculosis. 2012, 34(8):  523-526. 

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Objective To clone and express the genes Rv3803c and Rv3846 from Mycobacterial tuberculosis in Escherichia coli cells, and to purify Rv3803c and Rv3846 proteins. Methods The genes Rv3803c and Rv3846 were amplified from Mycobacterial tuberculosis H37Rv genomic DNA by polymerase chain reaction (PCR). PCR products were cloned into the expression vector pGEX-6P-1. After the plasmids were sequenced and validated their correctness, the recombinant plasmids were transformed into E. coli Rosetta strain to induce the expression of proteins. The proteins were purified by the GST-tag affinity chromatography, and then electrophoresed, stained with Coomassic brilliant blue, identified by Western-blot. Results The sequences of recombinant plasmids pGEX-Rv3803c and pGEX-Rv3846 are consistent with the design. The proteins GST-Rv3803c and GST-Rv3846 existed 50% in the soluble fraction and 50% in the inclusion body of E. coli Rosetta. Their molecular weights were 56 000 and 45 000, respectively. The purities of final products were higher than 90%. The concentrations of the purified recombinant proteins were approximately 0.1 mg/ml and 0.5 mg/ml, respectively. The yields of the recombinant proteins GST-Rv3803c and GST-Rv3846 were 0.25 mg and 1.25 mg from per liter of the E. coli Rosetta cultures, respectively. Conclusion The purified proteins Rv3803c（MPT51）and Rv3846（SodA）were successfully obtained, which provide the experimental basis for the further study of auxiliary diagnostic value of MPT51 and SodA proteins and construction of protective vaccines.

The practical evaluation of second-line anti-tuberculosis drug susceptibility test using MGIT 960 system

JIANG Jun, ZHANG Juan，ZHANG Hong, JIA Lin, WEI Cheng-cui, SHI Lian, MA Jing-hong, YANG Li-jun

Chinese Journal of Antituberculosis. 2012, 34(8):  527-531. 

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Objective  To evaluate the practicality of automatic BACTEC MGIT960 system in second-line anti-tuberculosis (TB) drug susceptibility test.  Methods 212 M. tuberculosis strains were isolated and idenfied from the sputum specimen in 435 cases by the smear, Lowenstein-Jenson (L-J) solid culture and MGIT960 liquid culture and identification. These strains were determined the resistance to first-line anti-TB drugs including isoniazid (INH), rifampicin (RFP),streptomycin (S) and ethambutol (EMB) by L-J proportion method, respectively. Then the first-line anti-TB drug-resistant strains were further determined the resistance to second-line anti-TB drugs including capreomycin (Cm), kanamycin (Km),ofloxacin (Ofx) and ethionamide (Eto)  by L-J and MGIT960, respectively.  The detection time and accordance rate of the two methods were compared.  Results The times of drug susceptibility test by MGIT960 and L-J method were average 10.0 days (1183/118) and 28.5 days (3360/118), respectively. There was difference of 18.5 days (28.5-10.0) between two methods. The accordance rates of MGIT960 and L-J method in Cm, Km, Ofx and Eto susceptibility test were 88.1%(104/118), 95.8%(113/118), 91.5%(108/118) and 71.2%(84/118).   Conclusion The second-line anti-TB drug susceptibility test using MGIT960 system is fast, accurate and has higher practical value.

Rpf-DNA vaccine for M. tuberculosis infected mice

XING Ai-ying, LIU Zhong-quan, LIU Yang, JIA Hong-yan, LI Zi-hui, ZHENG Xiao-jing,CAO Ting-ming, DU Feng-jiao, DU Bo-ping,GU Shu-xiang,ZHANG Zong-de

Chinese Journal of Antituberculosis. 2012, 34(8):  532-537. 

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Objective  To evaluate the immune protective effect of resuscitation promoting factor (Rpf) DNA vaccine on M. tuberculosis (Mtb) infected mice. Method  Rpf D and Rpf E plasmid DNA vaccines were construct. 180 BALB/c mice with 4 weeks of age were divided into 6 groups, 30 mice each group were immunized with empty plasmid, Rpf D plasmid DNA, Rpf E plasmid DNA, BCG and saline, respectively. All of mice were infected by aerosol with Mtb H37Rv. Serum anti-Rpf D or E antibodies and IFN-γ were detected. The lymphocyte proliferation assay and cytotoxicity test were performed, IFN-γ, IL-12 and IL-2 levels were tested after immunization with RpfD,E DNA vaccine. Histopathologic examination of the lung and CFU count were performed.  Results  (1) The antibody levels against RpfD and E were as follow：Rpf D group 0.32±0.1，Rpf E group 0.39±0.1,BCG group 0.02±0.01，empty plasmid group 0.01±0.0，saline group 0.09±0.04，blank group 0.03±0.01.The antibody levels in RpfD group and E group were significantly higher than those in BCG group，empty plasmid group,  saline group, and blank group (F value: Rpf D：（45.6，43.2，45.1，45.7）；Rpf E：（51.6，53.6，51.0，52.2）; P<0.01).  Serum IFN-γ levels: Rpf D group (43.9±24.8）pg/ml，Rpf E group (45.9±21.0）pg/ml,BCG group (21.0±11.0）pg/ml，empty plasmid group (7.9±4.9）pg/ml，Saline group (4.7±2.1）pg/ml，blank group (5.8±4.7）pg/ml，Serum IFN-γ levels of RpfD group and E group were higher than empty plasmid group, saline group, and blank group (F value: Rpf D：15.6，17.8，17.3；Serum IFN-γ levels of Rpf E：17.5，21.1，20.7; P<0.01). (2) The lymphocyte proliferation assay using cell counting kit-8 (CCK-8) showed: Rpf D group 176.0±4.2，Rpf E group 183.0±4.3,BCG group 101.0±1.1，empty plasmid group 100.0±6.8，saline group 104.0±8.4，blank group 116.0±2.2, which in vaccine groups were significantly higher than the other controls (F value: Rpf D：9.5，10.1，10.0，9.0；Rpf E：11.2，12.9，11.7，10.3，P<0.05). Cytotoxicity test showed: Rpf D group 32.0±3.2，Rpf E group 30.0±4.2,BCG group 16.0±5.9，empty plasmid group 3.3±1.5，saline group 6.7±0.5，blank group 7.3±3.5. The proliferation index in RpfD group and RpfE group was higher than the other control groups (F value Rpf D：8.8，14.5，13.7，11.9；Rpf E：8.1，12.5，11.6，11.1; P<0.05). (3) Cytokines in the supernatant of lymphocytes stimulated by Rpf D and Rpf E:IL-2 level: Rpf D group 9.5±2.4，Rpf E group 9.2±1.2,BCG group 2.4±2.1，empty plasmid group 1.2±0.3，saline group 1.8±1.0，blank group 1.5±0.7. The IL-2 levels in RpfD group and RpfE group were higher than the other control groups (F value Rpf D：12.5，14.6，13.5，13.9；Rpf E：12.0，13.6，13.1，13.2; P<0.05). IFN-γ level: Rpf D group 22.2±5.7，Rpf E group 28.7±14.4,BCG group 16.1±10.1，empty plasmid group 9.8±1.6，saline group 13.2±2.1，blank group 15.7±2.9. The IFN-γ level in RpfD group and RpfE group were higher than the other control groups（F value Rpf D：7.8，12.6，8.7，8.3；Rpf E：11.3，16.4，14.7，14.2; P<0.05）. Conclusion  RpfD and RpfE DNA vaccines can induce immune responses including humoral and cellular immunogenicity in Mtb infected BALB/c mice. Rpf might be used as the vaccine candidate.

Analysis of drug resistances and strain types in 214 Mycobacterium tuberculosis isolates in Kashi region, Xinjiang

Mireban·Rexiati，HU Xin，XU Yang-ling，WANG Jing，Muyesha’er·Bidayi，Tunishaguli·Taiwaikuli，JI Ping，Qimanguli·Wushou’er

Chinese Journal of Antituberculosis. 2012, 34(8):  538-541. 

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Objective  To analyze drug resistances to 8 kinds of anti-TB drugs and strain types in Mycobacterium tuberculosis clinical isolates in Kashi region, Xinjiang.   Methods  214 mycobacterial strains were cultured and isolated from sputum specimen from December 31, 2010, to June 1, 2010. and then identified their strain types and drug resistances to 8 kinds of anti-TB drugs, including isoniazid, rifampicin, streptomycin, ethambutol, ofloxacin, capreomycin, kanamycin, amikacin, by the proportional method for drug susceptibility testing. The count data between the two groups were compared using the χ² test, P<0.05 was considered as statistically significant difference.   Results  Of 214 isolates, 175 (81.78%) were Mycobacterium tuberculosis, 39 (18.22%) were Mycobacterium bovis. The drug-resistant rates had the statistical difference between Mycobacterium tuberculosis（29.14%，51/175）and Mycobacterium bovis （12.82%，5/39）(χ²=4.398，P=0.034). The total drug-resistant rate was 26.17%, initial drug-resistant rate was 25.97%（20/77）, and acquired drug-resistant rate was 26.28%（36/137）. They had not statistical difference (χ²=0.0003，P=1.00). The rates of resistance to 8 kinds of anti-TB drugs accounting for the top four were as follows: isoniazid, streptomycin, rifampicin, ethambutol.   Conclusion  The situation of drug resistance in TB patients is severe in Kashi region, Xinjiang. The infection of Mycobacterium tuberculosis is dominant.

The performance analysis of indicating growth of Mycobacterium tuberculosis complex by culture filtrate MPT64 antigen detection method

XU Wan-hua, HUANG Ye-lun, LIU Yan-wen, HU Li-huan, LUO Shao-zhen,MENG Fan-rong,LIU Zhi-hui

Chinese Journal of Antituberculosis. 2012, 34(8):  542-545. 

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Objective  To study the performance of indicating the growth of Mycobacterium tuberculosis complex (MTBC) by culture filtrate MPT64 antigen detection method.   Methods  799 BACTEC MGIT 960 mycobacterial cultures at 42 days and 1265 cultures every 7 days until the occurrence of a positive test result or the forty-second days were detected by MPT64 antigen detection method. Compared with BACTEC MGIT 960 mycobacterial culture system, the accuracy and rapidity of MPT64 antigen detection method were analyzed in 2064 BACTEC MGIT 960 cultures.   Results   (1) Of 2064 clinical samples, 298 MTBC strains, 108 non-tuberculous mycobacteria (NTM) strains, 1521 culture-negative cases and 137 cases of pollution were reported respectively by BACTEC MGIT 960 mycobacterial culture system, in which 279, 6, 0 and 2 samples were MPT64-positive respectively. The overall consistency of two methods was 98.69%（2037/2064）, The positive rate of MPT64 antigen detection method was 93.62%（279/298）in BACTEC-positive cultures. (2) Of 1265 cultures, 168 were identified as MTBC by these two methods. For these 168 samples, the positive rates of MTBC at 1 to 6 weeks were 25.60% (43/168), 66.67% (112/168), 92.26% (155/168), 98.21% (165/168), 98.81% (166/168) and 100.00% (168/168) respectively by BACTEC MGIT 960 mycobacterial culture system, while 17.26% (29/168), 61.90% (104/168), 89.88% (151/168), 95.24% (160/168), 98.81% (166/168) and 100.00% (168/168) respectively by culture filtrate MPT64 antigen detection method. The times of positive reports had not significant difference between two methods（t=1.68，P=0.366）.   Conclusion  The culture filtrate MPT64 antigen detection method to indicate growth of MTBC is accurate, fast, easy to operate and worthy of promotion in the clinical application. Especially, it is very suitable to primary hospitals without specific equipment and reagent.

The quality control of production of tuberculosis antibody detection reagents and the overview of its clinical research

CHEN Bao-wen，DU Wei-xin，YANG Lei，WANG Guo-zhi

Chinese Journal of Antituberculosis. 2012, 34(8):  546-549. 

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According to the guiding principle of the study and quality control on in vitro diagnostic reagents and “the registered management approach of in vitro diagnostic reagents” (for trial implementation) from State Food and Drug Administration, “the standard of diagnosis of pulmonary tuberculosis” and “the test procedures of tuberculosis diagnostic laboratory”, this paper explores the raw and auxiliary materials, production process, quality control of tuberculosis antibody detection reagents by colloidal gold method and ELISA, and the basic requirements of its clinical evaluation to provide the reference for the production and research units of tuberculosis serological diagnostic reagents and their clinical evaluation units.