Table of Content

10 September 2013, Volume 35 Issue 9

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Strengthen the scientific research on tuberculosis

PANG Yu, ZHAO Yan-lin

Chinese Journal of Antituberculosis. 2013, 35(9):  639-642. 

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The value of Mycobacterium tuberculosis fusion antigen 38F and 64F in the diagnosis of active tuberculosis

QIE Shuang，ZHANG Li, DAI Zhen-hua，ZHAO Ping，YANG Xi-qin，GUO Lan-qin，XIU Bing-shui, CHEN Kun, WANG Guo-hua, ZHANG He-qiu, FENG Xiao-yan

Chinese Journal of Antituberculosis. 2013, 35(9):  643-648. 

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Objective To explore the diagnostic value of Mycobacterium tuberculosis fusion antigen 38F and 64F in detecting IgG, IgM and IgA antibody in serum of patients with active pulmonary tuberculosis. Methods By using enzyme-linked immuno sorbent assay (ELISA) method, IgG, IgM and IgA antibody levels in serum of 223 patients with active pulmonary tuberculosis were detected with the Mycobacterium tuberculosis seven immune-dominat peptide fusion antigen 38F and 64F. Patients with active pulmonary tuberculosis were divided into three groups. The first group was co-positive cases of smear and culture examination (n=86). The second group was smear-negative and culture-positive (n=51). The third group was co-negative cases of smear and culture examination (n=86). Results The joint detection rate of IgG, IgM and IgA was 79.07% （68/86） for the first group, 62.75% （32/51） for the second group and 69.77% （60/86） for the third group. The detection rate of all samples by serological method was 71.75% （160/223）. Conclusion Mycobacterium tuberculosis fusion antigen 38F and 64F have high assistant diagnostic values to active pulmonary tuberculosis, and the detection rate is increased significantly through joint detection of IgG, IgM and IgA antibody.

Molecular characteristics of the second-line antituberculosis drug-resistant Mycobacterium tuberculosis strains from Jiangxi province

YUAN Xiao-liang, ZHANG Tian-tuo, ZHU Jia-xin, ZHENG Wen-zheng, LEI Jian-ping, TU Shao-hua, LUO Yi-jun, LIU Wei-you

Chinese Journal of Antituberculosis. 2013, 35(9):  649-654. 

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Objective To initially ascertain molecular characteristics of the second-line antituberculosis drug-resistant M. tuberculosis clinical isolates in Jiangxi, and to evaluate the feasibility of multiplex allele-specific polymerase chain reaction (MAS-PCR) assay for the detection of the second-line drug resistance in M. tuberculosis clinical isolates.  Methods Fifty-two second-line drug-resistant isolates and 30 pan-susceptible isolates from Jiangxi province were analyzed for gene mutations related to the second-line drug resistance using MAS-PCR and DNA sequencing.  Results DNA sequencing indicated that 32 of 39 ofloxacin （Ofx）-resistant isolates displayed missense mutations in gyrA gene, and the most common mutations were observed at codon 94 (n=27)，and only 2 isolates showed single-point mutation in gyrB gene. In addition, 22 of 29 kanamycin (Km)- or capreomycin (Cm)-resistant isolates had an A-to-G transition at nucleotide position 1401 of rrs gene. It is shown that the Beijing genotype was predominant (76.92%, 40/52) among the 52 drug-resistant strains. Of 40 Beijing genotype strains, 17 strains （42.50%）displayed gyrA-GAC94GGC mutations, and 19 strains (47.50%) showed rrs-A1401G mutations. No relationship could be demonstrated between Beijing genotype and gyrA-GAC94GGC or rrs-A1401G mutations（χ²=1.16，1.92，P value＞0.05）. In comparison with the phenotypic data, the sensitivities of the detection of Ofx resistance, and Km or Cm resistance using MAS-PCR were 61.54%（24/39）and 79.31%（23/29） respectively. Conclusion In Jiangxi province, gyrA mutation is the main molecular mechanism of Ofx resistance in M. tuberculosis, and an A1401G mutation in the rrs gene is the main cause of Km or Cm resistance. MAS-PCR, an assay with technical simplicity, short turnaround time, and low cost, could be useful for screening second-line drug-resistant M. tuberculosis, particularly in resource-limited areas with a high prevalence of tuberculosis and drug resistance.

Genotyping and drug resistance analysis of Mycobacterium tuberculosis in Ningxia

WANG Xiao-ping，PANG Yu，ZHAO Xiao，WANG Xiao-lin，WANG Yu-feng，XIAO Hui-xia，WANG Fu-rui，LIU Xiao-ni，ZHAO Yan-lin

Chinese Journal of Antituberculosis. 2013, 35(9):  655-659. 

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Objective To study the relationship between genotype and drug resistance of M. tuberculosis representative strains in Ningxia, and to investigate their molecular epidemiological characteristics. Methods One hundred and fifteen M. tuberculosis clinical isolates were collected in Ningxia, and analyzed their genotypes by spoligotyping and drug susceptibility by the proportional method. The chi-square test was used to compare the differences among the results of different groups. Results Of 115 M. tuberculosis isolates, 90 (78.3％) isolates belonged to Beijing genotype, 25 (21.7%) isolates belonged to non-Beijing genotype. Of 90 Beijing genotype isolates, 41(45.6%) isolates belonged to the ancient genotype, in which drug-resistant rate was 17.8% (16/90); 49 (54.4%) isolates belonged to the modern genotype, in which drug-resistant rate was 36.7% (33/90). There were no significant difference of drug resistances between different genotypes（χ²=1.27，P>0.05). Conclusion M. tuberculosis Beijing genotype was the predominant in Ningxia. There was no significant relationship between Beijing genotype and drug resistance.

The characteristic analyses in genotype and drug resistance of clinical isolates from the patients with tuberculous meningitis

WANG Ting, ZHAO Yan-lin, LIU Jia-yun, PANG Yu, ZHOU Yang, ZHAO Bing, ZHAO Gang

Chinese Journal of Antituberculosis. 2013, 35(9):  660-667. 

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Objective To study the characteristics in genotype and drug resistance of clinical isolates from the patients with tuberculous meningitis. Methods Twenty-five clinical isolates from the patients with tuberculous meningitis (TBM) were analyzed by Spoligotyping, variable numbers of tandem repeats (VNTR), 15 antituberculosis drug susceptibility testing methods, in which 15 drugs were as following: isoniazid, rifampicin, ethambutol, pyrazinamide, streptomycin, kanamycin, amikacin, capreomycin, moxifloxacin, ofloxacin, levofloxacin, para amino salicylic acid, ethionamide, prothionamide, cycloserine. Results Of 25 M. tuberculosis isolates, 20 (80.0%) were Beijing genotype, 2 were T1 genotype, 3 were other genotypes (including T2, LAM6, and unclassified genotype). 12.0%（3/25）were multi-drug resistant (MDR), and 4.0% (1/25) strains were extensive drug-resistant (XDR). 8.0% (2/25) strains were resistant to fluoroquinolones, all belong to Beijing genotype. 48.0% (12/25) strains were resistant to any drug, in which 83.3%（10/12）were Beijing genotype; 52.0%(13/25) strains were sensitive, in which 76.9% (10/13) strains were Beijing genotype. Conclusion Beijing genotype strains were possibly associated with drug-resistant tuberculosis and may act as the main epidemic strain of TBM. Few strains were resistant to fluoroquinolones, which may play an important role in TBM treatment.

Genotyping and drug resistance analysis of M. tuberculosis clinical isolates in Chongqing

WANG Xiao-yan, ZHAO Yan-lin, PANG Yu, WANG Yu-feng, WAN Kang-lin, LIU Jie, LIU Ying,SHEN Jing, DU Chang-ting, ZHANG Shun

Chinese Journal of Antituberculosis. 2013, 35(9):  668-672. 

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Objective To study the prevalence of different genotypes of M. tuberculosis in Chongqing and analyze the relationships between genotype and drug-resistant phenotype.  Methods One hundred and thirteen M. tuberculosis clinical isolates were collected in Chongqing from January to December in 2011, performed drug susceptibility testing by the proportion method, analyzed the genotypes by Spoligotyping and PCR method, and then made gene glustering analysis. Results One hundred and thirteen clinical strains were divided into 2 gene clusters, namely Beijing family (68.1%, 77/113) and non-Beijing family (31.9%, 36/113). Of 77 Beijing family strains, 48 (62.3%) strains were belong to modern type, 29 (37.7%) were the ancient type. Of 36 strains with non-Beijing family, 13 (36.1%) strains were T1 type, and found 9 new genotypes. The drug-resistant rate of Beijing family strains was 24.7% (19/77), and of non-Beijing family strains was 38.9% (14/36), in which there was no significant differences (χ²=2.40, P>0.05). The drug-resistant rate of modern type strains in Beijing family was 27.1% (13/48), and of the ancient type strains was 20.7% (6/29), in which there was no significant differences (χ²=0.39, P>0.05).  Conclusion There was obvious gene polymorphism in M. tuberculosis isolates in Chongqing. Beijing family was the main epidemic strains, in which the modern type was predominant. There was no significant correlation between genotype and drug resistance.

Anti-tuberculosis drug resistance analysis of patients with smear positive pulmonary tuberculosis in Guangxi

LAN Ru-shu, LAN Lan, HUANG Shu-hai, LIU Fei-ying, QU Jin, ZHANG Ying-kun, ZHAO Jin-ming

Chinese Journal of Antituberculosis. 2013, 35(9):  673-678. 

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Objective To investigate the drug resistant spectrum of patients with smear positive pulmonary tuberculosis (PTB) in Guangxi, and to clarify the distribution and law of drug resistance. Methods A prospective survey was conducted to recruit smear-positive PTB patients in the clinics of 30 TB control institutes in Guangxi. Sputum samples from 1765 cases were cultured and 1545 isolates of Mycobacterium tuberculosis were identified. Anti-TB drug susceptibility tests were conducted for 1545 isolates of Mycobacterium tuberculosis by conventional proportion method for isoniazide (H), rifampicin (R), ethambutol (E), streptomycin (S), ofloxacin (Ofx) and kanamycin (Km). Data for prior administration of anti-TB drugs were collected. Results The drug resistance rates of new cases to first line drugs was found in isoniazide 7.93% (94/1186), followed by streptomycin 6.24% (74/1186), rifampicin 3.20% (38/1186) and ethambutol 1.85% (22/1186), and those were 25.91% (93/359) for isoniazide, followed by rifampicin 25.07% (90/359), ethambutol 13.65% (49/359) and streptomycin 11.98%（43/359） in previously treated cases. The difference of proportion of any first line drug resistance between new cases and previously treated cases was statistically significant (χ²=43.174, P<0.01).There were 25 types of combinations of drug resistance among drug resistant cases. In the combinations to first line drugs, HRS (8 cases),HRE (5 cases) and HR (cases) were most common in new cases, and HRE (17 cases) and HR (15 cases) were most common in previously treated cases. Besides first line drug resistance, second line drug resistance was found in both new and previously treated patients. One cases was resistant to Km and 12 cases were resistant to Ofx in new cases. While in previously treated cases, the numbers were 6 and 31. Conclusion Results suggest that the orders of drug resistance rate of new and previously treated cases to first line drugs are different in Guangxi. There were 25 types of combinations of drug resistance among drug resistant cases, HRS and HR are most common in new cases, while HRE and HR are most common in previously treated cases.

Construction and immunogenicity of the DNA vaccine of dormancy related antigen Rv1733c of Mycobacterium tuberculosis

DUAN An-hu， ZHANG Wei， BAI Yin-lan， KANG Jian， WANG Rui，XU Zhi-kai， WANG Li-mei

Chinese Journal of Antituberculosis. 2013, 35(9):  679-685. 

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Objective To construct DNA vaccine of dormancy related antigen Rv1733c of Mycobacterium tuberculosis (Mtb), and to evaluate its immunogenicity in mice.  Methods Rv1733c gene was cloned into the eukaryotic expression vector pcDNA 3.1(-) from the plamid pMD-18T-Rv1733c, which was constructed previously and preserved in our lab. The constructed plasmid was named pcDNA-Rv1733c. P815 cells were stably transfected with the plasmid pcDNA-Rv1733c, and were detected the protein expression of Rv1733c by indirect immunofluorescence(IFT). The mice BALB/c were divided randomly into three groups named pcDNA-Rv1733c DNA, saline and BCG, 10 mice per group. The mice were immunized with pcDNA-Rv1733c DNA or saline intramuscularly 3 times at an interval of 2 weeks. BCG was immunized subcutaneously only once. The antigen specific antibody level and IgG2a/IgG1 subtype ratio in immunized mice were detected by ELISA every 2 weeks. At 8 weeks after the first immunization, the specific proliferation of spleno-lymphocytes of mice was detected by MTS ［3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-etrazolium, inner salt］ method, and IFN-γ secreted by spleno-lymphocytes was detected by ELISPOT. The percentages of CD4⁺ and CD8⁺ T cells in spleno-lymphocytes were analyzed by flow cytometry, and the function of cytotoxicity T lymphocyte (CTL) was detected by lactate dehydrogenase(LDH) assay． Results The eukaryotic expression plasmid of Rv1733c gene was successfully constructed, named pcDNA-Rv1733c. In P815 cells stably transfected with pcDNA-Rv1733c plasmid，the expression protein of Rv1733c could be stably detected by IFT. In immunized mice, the pcDNA-Rv1733c plasmid could stimulate the production of the antigen specific antibody, and the antibody subtype biased to IgG2a，but with the extension of immunization time，the ratio of IgG2a/IgG1 was tended to balance. In pcDNA-Rv1733c DNA group, the stimulation index of spleno-lymphocytes was (2.00±0.36), and the cell number of secreted IFN-γ was (41.48±5.30) SFC/10⁶, which were both statistically higher than those in saline group (t=3.096,P<0.05). However, the percentages of CD4⁺ and CD8⁺ T cells ［(18.15±2.30)% and (7.68±1.34)%］, and the activity of CTL ［(29.52±1.96)%］ were not significantly different with those in saline group(t=0.571, P>0.05).  Conclusion The eukaryotic expression plasmid of Rv1733c was constructed successfully. The pcDNA-Rv1733c plasmid DNA could induce specific humoral and cellular immunity in mice. It may be used as the new TB vaccine against latent Mtb infection.

Study on rapid differentiation of Mycobacterium tuberculosis complex from non-tuberculous mycobacteria by a multiplex PCR

WANG Gui-rong, FU Yu-hong, LIANG Qian, SHANG Yuan-yuan, JIANG Guang-lu, ZHAO Li-ping, YU Xia, CHEN Su-ting, HUANG Hai-rong

Chinese Journal of Antituberculosis. 2013, 35(9):  686-689. 

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Objective To establish and evaluate a multiplex PCR technique to rapid differentiate Mycobacterium tuberculosis complex (MTBC) from non-tuberculous mycobacteria (NTM). Methods Three pairs of oligo nucleotide primers were used in the multiplex PCR reaction. A 473 bp DNA fragment encoding oxyR-ahpC intergenic region in MTBC, a 235 bp and 136 bp DNA fragment encoding variable rpoB gene region from MTBC and NTM, respectively, were amplified. The multiplex PCR was assessed in 6 reference strains of MTBC, 50 reference strains of NTM, 312 clinical strains of MTBC and 300 clinical strains of NTM. Results The multiplex PCR produced two DNA fragments at the size 473 bp and 235 bp for MTBC reference strains, and one DNA fragment with the size 136 bp for NTM reference strains. Among 312 MTBC clinical samples, the sensitivity was 99.36% (310/312) and specificity was 99.32% (294/296). Among 300 NTM clinical samples, the sensitivity and specificity were 98.00% (294/300) and 100.00% (310/310) respectively. Conclusion The multiplex PCR can differentiate MTBC and NTM efficiently, and might be a valuable technique for clinical use.

Analyses of 23S rRNAs of 39 clarithomycin-resistant  Mycobacterium tuberculosis clinical isolates

KUANG Xiao-jia, TAN Shou-yong, WU Bi-tong, CAI Xing-shan, ZHANG Yuan-liang

Chinese Journal of Antituberculosis. 2013, 35(9):  690-692. 

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Objective To investigate the changes of 23S rRNA at site 2058 of clarithromycin-resistant Mycobacterium tuberculosis clinical isolates. Methods One Mycobacterium tuberculosis H37Rv standard strain and 64 isolates were detected, in which 39 clarithromycin-resistant isolates, 25 clarithromycin-sensitive isolates. The 23S rRNAs of Mycobacterium tuberculosis isolates were analyzed by polymerase chain reaction (PCR) and sequencing. Results The 23S rRNA of Mycobacterium tuberculosis H37Rv standard strain did not found the mutation at site 2058. Of 64 Mycobacterium tuberculosis isolates, only 1 extensively drug-resistant (XDR) strain had A2058A-G mutation of 23S rRNA, which account 2.56%（1/39） clarithromycin-resistant strains and 1/10 XDR strains. Conclusion The mutation of 23S rRNA at site 2058 of Mycobacterium tuberculosis may not be the main cause inducing its clarithromycin resistance. The mechanism of Mycobacterium tuberculosis clarithromycin resistance need be further studied.

The application research of tuberculosis specific antigen（TB-SA）antibody detection kit in the diagnosis of tuberculosis

LI Qiang, ZHAO Bing, XIA Hui, OU Xi-chao, FU Yan-yong, WANG Xie-xiu, WANG Hai-ying, PENG Lin, LI Hui, ZHAO Yan-lin

Chinese Journal of Antituberculosis. 2013, 35(9):  693-696. 

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Objective To evaluate the value of tuberculosis specific antigen (TB-SA) antibody detection kit in the diagnosis of tuberculosis. Methods Nine hundred and forty-four pulmonary tuberculosis suspects were continually recruited from April 2012 to October 2012 in 3 district’s TB centers from Tianjin city, Shandong province and Henan province, respectively，110 healthy people were enrolled at the same time. Sputum and serum specimen from all cases were collected for Ziehl-Neelsen staining smear, traditional mycobacterium culture and TB-SA antibody test. PPD skin test and chest X-ray were also examined to all cases. The data were entered separately by 2 people in sites and analyzed by national TB reference laboratory. Results Among 755 active TB patients, the positive rate of TB-SA antibody test were 74.8% (565/755, 95%CI: 71.7%-77.9%), significantly higher than smear (25.6%, 193/755，χ²=366.59, P<0.0001) and culture (39.6%, 299/755, χ²=190.12，P<0.0001); its specificity and positive predictive value were 97.3% (95%CI: 94.3%-100.3%) and 92.3% (95%CI: 90.2%-94.4%), respectively. Among culture-positive TB patients, the sensitivity of TB-SA antibody test was 88.5% (95%CI: 84.8%-92.2%). However, the sensitivity of TB-SA antibody test was 68.0% (95%CI: 63.7%-72.3%) among smear- and culture-negative TB patients. Conclusion TB-SA antibody test has significantly higher positive rate than smear and conventional culture, and can be used for the diagnosis of TB patient especially smear- and culture-negative TB patients. In addition, it can be used for healthy screening examination.

Expression patterns of interleukin-8 during the progress of disease in 40 pulmonary tuberculosis patients

CHE Nan-ying, LI Xue-lian, ZHANG Xu-xia, GAO Meng-qiu, LI Chuan-you

Chinese Journal of Antituberculosis. 2013, 35(9):  697-701. 

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Objective To study the expression patterns of interleukin-8 (IL-8) in peripheral blood samples from 40 pulmonary tuberculosis (PTB) patients during progress of PTB.  Methods Fifteen healthy donors and 40 newly diagnosed PTB patients were recruited from May 2011 to February 2013 in Beijing Chest Hospital. The PTB patients were divided into 3 groups: 8 patients for tracking studies; 17 patiens with mild symptoms (negative sputum bacteria, the lung without cavity); 15 patients with severe symptoms (positive sputum bacteria, the lung with cavity). Anticoagulant peripheral blood samples from 8 patients for tracking studies were collected before treatment, during treatment (2 to 3 months after treatment started), and after treatment (1 to 3 months after treatment ended), while that from the other 32 patients were collected only before treatment. Then the blood samples were incubated with Mycobacterium tuberculosis whole cell proteins. Plasma IL-8 expression levels were detected by enzyme linked immunosorbent assay. Statistical analysis was performed using SPSS 17.0 software. Mann-Whitney U test was used for analyzing IL-8 expression differences among healthy donors, mild PTB patients, and severe PTB patients. Wilcoxon test was used for comparing IL-8 expression levels before, during, and after treatment. IL-8 value was shown as (x±s), P value<0.05 was considered as statistically significant.  Results Induced expression level of IL-8 in severe PTB patients was (176.7±133.2) ng/ml, while that in mild PTB patients was (39.6±52.2) ng/ml. Both expression levels were significantly higher than that in healthy donors ［(7.1±6.3) ng/ml］(U=5, P=0.000 008; U=62, P=0.013). Also, expression level of IL-8 in severe PTB patients was significantly higher than that in mild PTB patients (U=37, P=0.001). Additionally, IL-8 expression level in patients after treatment ［(5.9±3.6) ng/ml］was significantly decreased than those before treatment ［(34.9±33.7) ng/ml］and during treatment ［(51.7±34.2) ng/ml］(Z=－2.240, P=0.025; Z=－2.521, P=0.012). Conclusion IL-8 expression level is associated with progress of PTB.

Clinical value of endotoxin and C-reactive protein on the diagnosis of pulmonary tuberculosis patients with gram negative bacteria

LI Cheng-hang，LI Yue-cui

Chinese Journal of Antituberculosis. 2013, 35(9):  702-705. 

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Objective To explore the clinical value of endotoxin and C-reactive protein (CRP) on tuberculosis (TB) patients with gram negative bacteria. Methods Two hundred and fourty TB inpatients were analyzed by retrospective investigation. According to the results of sputum culture and hemoculture, the patients were divided into gram-negative bacterium group, gram-positive bacterium group and culture-negative group. The concentrations of endotoxin and the levels of CRP in plasma from these patients were analyzed. Results The concentrations of endotoxin (317.34 pg/ml) and the levels of CRP (65.43 mg/L) in gram-negative bacterium group were significantly higher than that in gram-positive bacterium group (34.78 pg/ml, 35.33 mg/L) (Z=－1.793, Z=－1.106, P<0.05), and those in gram-negative bacterium group were significantly higher than that in culture-negative group (20.06 pg/ml, 15.34 mg/L) (Z=－2.397, Z=－1.950, P<0.01); There was correlation between endotoxin and CRP in gram-negative bacterium group (r=0.458, P<0.05). Conclusion The detection of endotoxin and CRP in TB patients will help to rapidly indentify gram-negative bacterium infection, improving the accuracy of clinical diagnosis and making reasonable prescription.

Evaluation of multidrug-resistant tuberculosis control in 5 years in Beijing

HONG Feng, GAO Zhi-dong, LI Bo, SUN Shan-hua, ZHAO Xin

Chinese Journal of Antituberculosis. 2013, 35(9):  706-710. 

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Objective To describe drug resistance spectrums of tuberculosis(TB) isolates from multidrug-resistant tuberculosis (MDR-TB) suspects, to assess the treatment effect of domestic drug group of MDR-TB and to explore the MDR-TB control strategy which is applicable in Beijing. Methods During the period of April 2008 to March 2013, 1124 MDR-TB suspects were detected by sputum culture in 21 651 patients with active tuberculosis. Drug susceptibility test (DST) was applied to the microbe-positive cultural substances of 885 MDR-TB suspects with strains of Mycobacterium tuberculosis complex, 277 diagnosed MDR-TB patients were treated by domestic drug prescription, and the treatment management methods were selected according to the patient’s situation, and the adverse effect and therapeutic effect were observed.  Results During the 5 years of implementation, MDR-TB patients accounted for 32.1%（284/885）of the MDR-TB suspects with Mycobacterium tuberculosis strains. The proportion of the strains which were sensitive to both the first and the second-line drugs were 21.4%(133/620). Because 7 MDR-TB patients conducted DST again during treatment, there were actually 277 MDR-TB patients detected. Classified management methods were applied to the 277 MDR-TB cases, 127 cases (45.9%) were treated with the second-line drugs in TB control institutes, among whom 76 cases(59.8%) had side effects. The cure rate of the 93 MDR-TB patients who had treatment outcomes were 63.4%(59/93). The proportions of MDR-TB patients maintained the original treatment regimen or referred to a specialized hospital were 17.7%(49/277) and 13.0%(36/277). Conclusion It is essential for the suspected MDR-TB to conduct DST timely to, get early detection of MDR-TB cases and to develop an appropriate treatment regimen to control the spread of multi-drug resistant TB. The classified management for MDR-TB patients gets the ideal effect.

Survey on drug resistant pulmonary tuberculosis and analysis of risk factors to drug resistance of retreatment cases in Guangxi

HUANG Shu-hai, LAN Ru-shu, LIU Fei-ying, ZHAO Jin-ming, LAN Lan, OU Jin, ZHANG Ying-kun

Chinese Journal of Antituberculosis. 2013, 35(9):  711-717. 

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Objective To explore the status of drug resistant pulmonary tuberculosis and analyze the risk factors contributing to drug resistance of retreatment patients in Guangxi. Methods A prospective survey was conducted by consecutively recruiting patients with smear-positive pulmonary tuberculosis (TB) in 30 TB control institutes from 2010 to 2011 in Guangxi. Forty-one new cases and 22 retreatment cases were assigned to be recruited in each institute. Information on patient’s socioeconomic status and previous clinical history was collected by questionnaire. Identification of Mycobacterium tuberculosis complex (MTBC) was based on para-nitrobenzoic acid culture test. Drug susceptibility tests (DST) were performed for isoniazid, rifampicin, ethambutol, streptomycin, kanamycin and ofloxacin. Univariate analysis was applied to each variable of patients’ socioeconomic and clinic information to assess the risk factors associated with drug resistance. Results One thousand five hundred and forty-five isolates collected in the survey were identified as MTBC. Among them, 17.22% (266/1545) cases were resistant to at least one anti-TB drugs, drug resistant rates of new smear positive cases and retreatment cases were 11.97% (142/1186) and 34.54% (124/359) respectively. The drug resistant rate of retreatment cases was higher than new cases (χ²=98.473, P=0.000). 6.28% (97/1545) cases were multidrug-resistant tuberculosis (MDR-TB), 2.45% (29/1186) for new cases and 68 (18.94%,68/359) for cases treated previously. MDR-TB rate was much higher in previous treated cases than new cases (χ²=127.450, P=0.000). 0.19% (3/1545) of cases was found to be extremely drug-resistant tuberculosis (XDR-TB), all of whom were retreatment cases (0.84%, 3/359). Univariate analysis of drug resistance of retreatment cases indicated that female (OR=2.009, 95%CI: 1.145-3.523, χ²=6.062, P=0.014), Zhuang nationality (OR=1.609, 95%CI: 1.024-2.529, χ²=4.289, P=0.038), selecting general hospital for first health care seeking (OR=1.967, 95%CI:1.210-3.198, χ²=7.565, P=0.006), more than once prior anti-TB treatment (OR=4.128, 95%CI:2.506-6.801, χ²=33.160, P=0.000), nonstandard treatment regimen (OR=3.419, 95%CI:1.952-5.988, χ²=19.775, P=0.000), and low family income (OR=4.777, 95%CI: 1.117-20.435, χ²=5.336, P=0.021) were found to contribute to higher trend of MDR-TB.  Conclusion Prevalence of drug resistant TB in Guangxi is lower than the national level. Results indicates that female, Zhuang nationality, low family income, seeking health care in non-designated institute and repeated treatment may be the risk factors of drug resistance.

Evaluation of genechip in detection of multidrug-resistant Mycobacterium tuberculosis in the prefectural-level laboratory

ZHAO Bing, SHI Jin-yan, PANG Yu, XIA Hui, LI Qiang, OU Xi-chao, SONG Yuan-yuan, WANG Yu-feng, CHI Jun-ying, ZHAO Yan-lin

Chinese Journal of Antituberculosis. 2013, 35(9):  718-722. 

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Objective To evaluate the performance of genechip in detecting rifampicin resistant, isoniazid resistant and multidrug-resistant tuberculosis (MDR-TB), and to analyze the feasibility of genechip application in the prefectural-level TB laboratory. Methods The sputum samples collected from 745 smear positive TB patients detected in Lianyungang city from January 1st 2011 to March 31st 2012 were applied by both genechip and conventional drug susceptibility testing (DST). Out of 745 TB patients, 111 were excluded due to negative culture result and non-tuberculous Mycobacteria. The conventional DST results were served as the gold standard, the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were analyzed among 634 TB patients. Results The results showed that the sensitivity and specificity of genechip were 84.4% (65/77) and 97.7% (544/557) for rifampicin resistance, 80.9% (76/94) and 97.4% (526/540) for isoniazid resistance and 70.0% (42/60) and 98.3% (564/574) for MDR respectively. The costs of genechip and conventional DST for detecting one smear positive TB patient were 165.65 yuan and 374.07 yuan,respectively. Conclusion Genechip is a more effectient, rapid, safer and more cost-effective diagnosis tool for drug-resistant tuberculosis in Chinese primary laboratories.

Cost analysis of Xpert Mtb/RIF test for the detection of M. tuberculosis and rifampin resistance

OU Xi-chao, XIA Hui, LI Qiang, PANG Yu, ZHAO Bing, ZHANG Zhi-ying, LI Jun-chen, ZHANG Jian-kang, CHI Jun-ying, ZHAO Yan-lin

Chinese Journal of Antituberculosis. 2013, 35(9):  723-728. 

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Objective To compare the cost of solid culture, drug susceptibility test and Xpert Mtb/RIF test in the primary laboratories in China. Methods We collected the cost data of solid culture for 3 times and Xpert Mtb/RIF test from 4 county (district) level laboratories, and strain identification and rifampin (RFP) susceptibility test for 3 times from 2 province reference laboratories. The element method was used to calculate the unit cost of each method, the detection cost of each patient. Of the population with different TB prevalence rate, the solid culture was used as the gold standard to compare the cost of Xpert Mtb/RIF test to find 1 TB patient under different detection efficiency. Of the population with different rifampicin-resistant prevalence rate, the traditional drug susceptibility test was used as the gold standard to compare the cost of Xpert Mtb/RIF test to find 1 RFP-resistant patient under different detection efficiency. After price adjustment of Xpert Mtb/RIF detection equipment and reagents, the unit cost of Xpert Mtb/RIF test was analyzed. Results The unit costs of solid culture, strain identification, RFP susceptibility test and Xpert Mtb/RIF detection were 47.87, 46.73, 82.86 and 118.62 Yuan, respectively. The cost per TB patient by traditional methods (2 solid cultures, 1 sub-culture, 1 PNB strain identification) was 172.57 Yuan. The average cost of each patient with drug-resistant TB by traditional methods (2 solid cultures, 1 sub-culture, 1 PNB strain identification and 1 drug susceptibility test) was 208.84 Yuan. When the specificity of Xpert Mtb/RIF test for the detection of Mtb and RFP was 85%, its sensitivity is greater than 70%, its cost of detecting one TB case or RFP-resistant case among the population with 1% to 70% prevalence rate of TB or RFP resistance was lower than traditional methods. The influence of reagent price changes on the testing cost was far greater than the change of equipment price. Conclusion The Xpert Mtb/RIF test is a rapid and more economic method than traditional tests, and suitable for the use in the primary laboratories.

Effect evaluation of Shenzhen tuberculosis control and prevention programme implementation from 2001 to 2010

GUAN Hong-yun, YANG Ying-zhou, TAN Wei-guo, WU Qing-fang, LV De-liang

Chinese Journal of Antituberculosis. 2013, 35(9):  729-737. 

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Objective To evaluate the implementation effect and the achievements of the objectives of Shenzhen tuberculosis control and prevention programme from 2001 to 2010，and to provide scientific recommendations for sustainable development of TB control. Methods The data from TB reporting system and TB Control and Prevention Programme final evaluation between 2001 to 2010 were collected, SPSS 13.0 was applied to analyze the financial investment、human resources and DOT coverage rate, referral and tracing situation of 195 201 TB suspects from 2005—2010, as well as the detection、treatment and management situation of 34 024 TB patients, and the epidemiology situation and social and economic benefits from 2001 to 2010. Results (1)Shenzhen implemented DOT strategy for the entire region，all people and all types of patients, the coverage rate reached 100.00%（660/660）.(2)The number of TB control staff in 2010 were 152, which was increased by 70.79%(63/89), 55.26% (84/152) of the staff had bachelor degree or above.(3) The overall arrival rate of the suspects among floating population was significantly lower than the resident population during 2005—2010, which were 69.21% (127 583/184 334) and 99.03% (10 762/10 867) respectively (χ²=4420.76，P＜0.01). (4) A total of 34 024 active pulmonary tuberculosis cases were detected from 2001 to 2010, among which 22 008 were smear positive cases and 12 016 were smear negative cases. The average cure rate of smear positive cases was 85.36% (18 787/22 008), and the average treatment success rate was 89.03% (30 292/34 024). The average cure rate and treatment success rate of the floating patients were 82.91%(14 479/17 463) and 87.25%(23 290/26 693), which was significantly lower than the resident patients (94.79%(4308/4545), 95.51%(7002/7331)) (χ²=406.92，401.91，P＜0.01). (5) The prevalence rate of smear positive of resident population was 19.44 per 100 000 population, which decreased 70.55% (46.56/66.00) compared with that of 2000, and that of the floating population was 59.96 per 100 000 population, which decreased 27.23% (22.44/82.40) compared with that of 2000.(6) About 0.15 billion yuan was invested in TB control from 2001 to 2010, 136 700 healthy people were protected from TB infection and 13 700 people were prevented to be new TB cases, the direct and indirect social and economic benefits reached 20.245 billion yuan. For every 1 yuan input into the TB control, 134.98 yuan of social and economic benefits can be produced. Conclusion Shenzhen has made substantial progress to control TB, while there are still some new challenges to be faced during the implementation of the Twelfth Five-year Plan.

Surgical therapy for 78 cases with tuberculous empyema

DOU Xue-jun, WANG Liang, LU Wei-qiang, WANG Zhi-yuan, MA Yun-lei, LI Liang

Chinese Journal of Antituberculosis. 2013, 35(9):  738-740. 

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Objective To investigate the outcome of patients with tuberculous empyema who underwent surgery. Methods We analyzed retrospectively the data from 78 patients with tuberculous empyema admitted and undergone surgical operation in the Aerospace Center Hospital and Beijing Chest Hospital during January 2007 and December 2012.  Results Postoperative complications occurred in 7 patients(9.0%)，including internal hemorrhage, wound infection and drug-induced hepatitis accompany with fluid and electrolyte imbalance in a different patient, they were recovered after treatment. In addition, respiratory failure in 4 patients, 3 of which were cured by treating with breathing machine and 1 patient died. Mortality was 1.3%.Seventy seven patients received antituberculous drug treatment for 6 months to 24 months. One patient suffered from pneumonia in six months and then recovered with anti-inflammatory therapy. After follow-up for 2 years, 77 patients had satisfactory efficacy.  Conclusion Surgical operation treating with tuberculous empyema would give the best chance of cure with less complications.

The clinical feature and treatment of pulmonary tuberculosis complicated with cor pulmonale in 140 Uygur young adults

SHANG Yong, HE Tao, Dilala TU’ERXUN, Shajidan YASHENG，CHEN Xiao-yu

Chinese Journal of Antituberculosis. 2013, 35(9):  741-743. 

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Objective To analyze the clinical feature and treatment of pulmonary tuberculosis (PTB) complicated with cor pulmonale in Uygur young adults in order to reduce misdiagnosis. Methods The course of disease, clinical feature and treatment of 140 Uygur young adults with PTB complicated with cor pulmonale aged 20-55 hospitalized in the First People’s Hospital of Kashi in 2010—2012 were analyzed retrospectively. Results Among the 140 cases, there were 28 cases with hemoptysis, 140 with pulmonary rales, 68 with emphysema sign, 81 with liver enlargement and leg edema and 45 with hydrothorax and seroperitoneum. Sixty-two cases were diagnosed with cor pulmonale by EGG, 60 with P pulmonale, 59 presented low voltage of QRS complex in the limb leads, 51 with arrhythmia. Seventy-eight cases were diagnosed with cor pulmonale by color doppler echocardiography. After treatment, 121 cases’ condition improved and 19 cases died. Conclusion As Uygur young adults with PTB complicated with cor pulmonale have atypical symptoms, measures should be implemented for timely detection and standardized treatment to reduce mortality.

Discussion on clinical trial, production and quality control of in vitro Mycobacterium tuberculosis IFN-γ detection reagents

DU Wei-xin, CHEN Bao-wen, XU Miao, YANG Lei, LU Jin-biao, WANG Guo-zhi

Chinese Journal of Antituberculosis. 2013, 35(9):  744-747. 

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According to Management Method of In Vitro Diagnostic Reagents Registration (Interim) and other relevant laws and regulations, we summarize respectively the main raw materials, production process, product quality control and technical requirements of clinical study for in vitro Mycobacterium tuberculosis IFN-γ detection kits and provide technical support for the development and registration for such kits.

Research progress on the detection methods of Mycobacterium tuberculosis gene mutation

LIU Zheng-wei, HUANG Yu, ZHAO Yan-lin

Chinese Journal of Antituberculosis. 2013, 35(9):  748-751. 

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The completion of the Mycobacterium tuberculosis genome sequencing provides the evidence for the study of genetic nature of tuberculosis bacterium, and the study of Mycobacterium tuberculosis gene mutations has become one of hotspots in the field of TB research at present. The development and application of new technology of molecular biology, such as high-resolution melting (HRM), gene chip, DNA direct sequencing have provided more effective methods for Mycobacterium tuberculosis gene mutation research. This paper reviewed relative literatures in recent years at home and abroad, introduced in category the principles and application of different methods in Mycobacterium tuberculosis gene mutation detection. Gene mutation detection technology such as GeneXpert, Mycobacterium tuberculosis linear probe (line probe assay, LiPA), gene chip, which are more mature and are expected to be widely used at present were introduced as well.

Research progress of control strategy on tuberculosis outbreak in school

LU Xi-wei

Chinese Journal of Antituberculosis. 2013, 35(9):  752-756. 

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Tuberculosis outbreak is one of serious public health challenges in schools in China. This article reviews challenges that tuberculosis control faces in school, field epidemiologic surveys and measures dealing with tuberculosis outbreaks. It summarizes particularly the research progress of screening tools and treatment of latent tuberculosis infection and concludes that early detection of pulmonary tuberculosis cases is the fundamental approach of tuberculosis outbreak control, combination of exposure with TST is an effective means to evaluate the latent tuberculosis infection, short-term chemotherapy and full administration are critical approaches to improve patient compliance. This article provides a reference for the development of scientific strategy on tuberculosis control in school.