Table of Content

10 July 2012, Volume 34 Issue 7

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Clinical application of loop-mediated isothermal amplification method in rapid detection of Mycobacterium tuberculosis

CHEN Tao, ZHOU Lin, ZHOU Jie, LI Hai-cheng, JIANG Yong, PENG Dong-dong, ZHOU Zhi-gang, PENG Jian-ming, WEN Li, HE Chao-wen, QIAN Ming, SUN Yi-fan, SHI Lei, ZHONG Qiu

Chinese Journal of Antituberculosis. 2012, 34(7):  413-418. 

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Objective  To evaluate the diagnostic value of a loop-mediated isothermal amplification (LAMP) method for rapid detection of Mycobacterium tuberculosis.  Methods  2129 sputum samples of suspected TB patients from 6 cities in Guangdong province and 337 enriched sputum samples randomly selected from Guangzhou and Shantou cities were collected. M. tuberculosis in 2129 sputum samples were detected by smear, Roche solid culture and LAMP methods. M. tuberculosis in 337 sputum samples were detected by quantitative polymerase chain reaction (qPCR) and LAMP methods.  Results  For 2129 sputum samples, the positive rates of smear, culture and LAMP were 13.2%（282/2129）, 21.7%（463/2129）and 20.3%(432/2129), respectively. Compared with Roche culture method, sensitivity, specificity, positive predictive value and negative predictive value of the LAMP method were 89.1%(432/485), 95.5%(1570/1644), 85.4%(432/506) and 96.7%(1570/1623), respectively. For 337 enriched sputum samples, the positive rates of qPCR and LAMP were 35.0% (118/337) and 35.9% (121/337), respectively. Compared LAMP with smear, culture and qPCR, the consistent rate of the results were 87.9%（K=0.612）, 96.1%（K=0.89）and 91.4%（K=0.812）, respectively. Conclusion  The LAMP method was equivalent to culture and qPCR, superior to smear method in detecting M. tuberculosis of sputum specimens. In addition, LAMP method showed the advantages of rapid, simple operation and no equipment, indicating good prospects for clinical application and promotion in the diagnosis of tuberculosis.

Operational feasibility of medication monitors in monitoring treatment adherence among TB patients

HUAN Shi-tong, CHEN Rong, LIU Xiao-qiu, OU Xi-chao, JIANG Shi-wen, ZHAO Yan-lin, ZHANG Zhi-ying, ZHAN Si-yan

Chinese Journal of Antituberculosis. 2012, 34(7):  419-424. 

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Objective  To explore the operational feasibility of using medication monitors to monitor treatment adherence in TB patients by comparing the medication monitors’ records with those of patients’ urine tests for traces of TB drugs.   Methods  Four hundred and thirty-two newly registered initial treatment TB patients were enrolled in succession in 9 counties of China MOH-Gates Foundation TB Control Project case management pilot sites in Heilongjiang, Hunan, Chongqing and Jiangsu provinces. The male-to-female ratio of the enrolled patients is 3.15:1, and average age is (45.98±17.36) years old. Among all the enrolled patients, 226 cases were in the intensive phase of treatment and 206 cases were in the continuous phase of treatment. All the patients were treated using the regimen 2H₃R₃Z₃E₃/4H₃R₃. An electronic medication monitor stored the patients’ drugs and kept a record of every time patients took their medicine out of the monitor. Investigators determined the urine test date for patients by using a table of random numbers at the patients’ 2nd or 4th month of treatment. Investigators then visited the patients’ house to collect patients’ urine and conducted a simple urine test to detect rifampicin to confirm whether patients had taken their medicine within the past 24 hours. Among the 319 patients tested, 116 cases were in the intensive phase of treatment and 153 cases were in the continuous phase, with an overall test proportion of 73.84% (319/432). Among the 113 patients not tested, 64.60% (73/113) were not at home on the test day, 20.35% (23/113) had changed their treatment regimen or quit due to adverse effects of drugs, and 8.85% (10/113) had taken their drugs less than 6 hours before the doctors’ visit and did not meet the criteria for the urine test, 4.42% (5/113) had taken their drugs more than 24 hours before the doctors’ visit and the doctors misclassified them as unqualified cases and did not conduct urine test, and 1.77%（2/113） refused to the testing. The operational feasibility of medication monitors in monitoring adherence to TB treatment was evaluated in terms of sensitivity and specificity by comparing 24h intake records from the monitor against rifampicin incidence in urine samples.  Results  Compared to the detection of rifampicin in urine, the sensitivity and specificity of 24h medication monitor records as the evidence of TB drug adherence were respectively 99.52% (208/209) (95% CI：97.30%-100.00%) and 95.45% (105/110) (95% CI：91.56%-99.35%). Meanwhile the sensitivity (Male: 99.38%(160/161)，95% CI：98.16%-100.00%；Female:100.00%(48/48)，95% CI：87.90%-100.00%）and specificity（Male：95.24%(80/84)，95% CI：90.68%-99.79%；Female: 96.15%(25/26)，95% CI:88.76%-100.00%）between gender groups and the sensitivity（Intensive phase:99.06%(105/106)，95% CI:97.21%-100.00%；Continuous phase：100.00%(103/103)，95% CI:93.00%-100.00%）and specificity（Intensive phase:93.33%(56/60)，95% CI：87.02%-99.65%；Continuous phase:98.00%(49/50)，95% CI：97.12%-100.00%）between treatment phases show no significant statistical difference (χ²=0.8，0.0，0.8，0.0，1.5；P>0.05).   Conclusion  There is a high correlation between medication monitors’ records and the TB treatment adherence by the patients; hence the records of medication monitors can be used as evidence of TB patient’s treatment adherence.

Risk factors of pulmonary tuberculosis incidence in Guangzhou: a paired case-control study

HE Zhi-qing，HU Gui-fang，ZI Qing-lan，TANG Hui-hong，GONG Fang，CHEN Shu-zhong，LEI Yu，BAO Wan-ling，LIU Gui-si

Chinese Journal of Antituberculosis. 2012, 34(7):  425-432. 

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Objective  To explore the risk factors affecting the TB incidence in Guangzhou, and provide scientific basis for pulmonary tuberculosis(PTB) prevention.  Methods  This is a 1：1 matched case-control study. Two hundreds and fifty smear positive tuberculosis patients in case group and the same number of people selected from household close contacts of smear positive in control group were investigated by unified self-designed questionnaire. A total of 25 indicators were surveyed including level of education, occupation, personal monthly income, the family economic responsibility, family economic status, smoking, excessive alcohol consumption, exercise, marriage, drug abuse, history of diabetes, the housing area, room ventilation, darkness and dampness, the surrounding environmental quality and 9 psychological stress indicators. The psychological stress was investigated by Yang Desen life Event scale (LES), Jiang Qianjin Trait Coping Style Questionnaire（TCSQ），Xiao Shuiyuan social support rating scale（SSRS）. Measured by the pre-survey, the Cronbach’s coefficient of the questionnaire was 0.83. Two hundred and sixty-eight pieces of questionnaire were issued for PTB patients, and 250 valid were returned with the effective response rate of 93.3%. All these 250 patients were enrolled in the case group. 1206 pieces of questionnaire were issued for household close contacts, and 1143 valid were returned with the effective response rate of 94.8%. Among these 1143 household contacts, 250 controls were selected according to 1：1 match with cases. The matching principle is same gender, and age gap less than 3 years old. The univariate analysis is made to screen the risk factors that may affect the TB incidence, and then the multivariate conditional logistic regression model is made for analysis. Wald χ², P values, OR values and their 95% CI is calculated for each indicator. P value ≤ 0.10 is considered statistically significant.  Results  A total of fifteen indicators are statistically significant in univariate analysis, nine enter into the multivariate analysis. Among which, six are risk factors of TB incidence. They are personal monthly income (three levels, <1000 yuan/month, 1000— yuan/month, ≥ 3000/month, Wald χ² value is 13.492,1.379, P=0.000,0.240), bearing the primary responsibility of family income  (Wald χ² =17.820, P=0.000), smoking (Wald χ² =9.489, P=0.002), excessive alcohol consumption (Wald χ²=4.141, P=0.042); the total amount of stimulation of the life events (Wald χ² =6.995, P=0.008), negative coping (Wald χ² =14.806, P=0.000. Their OR value (95% CI values) are 4.178 (1.948—8.961), 1.475 (0.771—2.821), 6.457 (2.716—15.352), 2.960 (1.484—5.905), 5.048 (1.062—24.004), 1.030 (1.008—1.052), 1.131 (1.062—1.204) respectively. Conclusion  Low personal incomes, bearing the primary responsibility for the family income, smoking, excessive alcohol consumption, life event stimulation and negative response are the risk factors of PTB incidence in Guangzhou.

The impact of TB core information on TB awareness among general public

GAO Cui-nan, XU Zhuo-wei, TAN Qing-yun, YANG Qiong, WU Jin-yao, ZHOU Lin

Chinese Journal of Antituberculosis. 2012, 34(7):  433-436. 

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Objective  To understand the impact of the numbers of TB core information on TB awareness in order to explore the most scientific and straightforward ways of expression, and meet the proposed 85% of TB awareness target set by National TB Control Program(2011—2015).  Methods The self-designed questionnaire was used in this survey. The people who pass through the gate of the hospital were sequentially investigated by health worker. 272 copies of questionnaire were distributed and 260 copies returned. The response rate is 95.6%.  Results Of the 260 copies of valid questionnaires, the awareness rate of first four pieces of TB Core Information were 84.7%(881/1040), 4.9 percent higher than that of all five pieces 79.8%（1037/1300）. The difference is statistically significant (χ²=9.55, P=0.00). In youth and old-age groups, the awareness rate of all five pieces was also significantly lower than that of the first four pieces, (85.0% and 90.5%, 71.1% and 77.4% respectively, χ²=4.58，4.29；P＜0.03、0.04). In terms of awareness rate, for the middle-aged group, although there has no statistically significant difference between four pieces and five pieces, the rate of four pieces is 3.1 percent higher than five pieces. In comparison of awareness rate of four pieces and five pieces in different education level, 77.4% and 67.1% for people with primary education, and 84.7% and 81.2% for people with secondary education, and 88.8% and 82.5% for people with college education, there is no significant difference (χ²＝3.62，3.56，3.60；P=0.06，0.06, 0.06).  Conclusion Adopting questionnaire with four pieces of TB core information can improve the TB awareness rate than that with five pieces of information. In public awareness survey, this could replace the questionnaire with five pieces of TB Core Information.

Clinical analysis of classic debridement surgery in the treatment of 104 patients with complex and compound spinal tuberculosis

CAO Ye, SONG Yan-zheng, LI Lei, SI Yan-hui, LIU Bao-chi, WANG Xu

Chinese Journal of Antituberculosis. 2012, 34(7):  437-440. 

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Objective  To explore the perioperative management, the surgical techniques and operation time of classic debridement surgery in the treatment of complex and compound spinal tuberculosis, in order to reduce the complications of these patients.  Methods  Clinical effects of 104 patients with complex and compound spinal tuberculosis of classic debridement surgery were retrospectively analyzed.  Results  The 104 patients had no death and serious complications with heart,lung or kidney. Among the 104 patients, 90 cases were performed for the first surgery, in which postoperative primary healing rate was 91%(82/90), the left 8 cases were healed after dressing and drainage, not for a second surgery on the same side. The 14 cases from the other hospital (referring to the third hospital) were healed. The lesions were removed through debridement in the opposite line, the sinus tract of the original knife-edge line were healed by resection. ESR of all the patients was normal after 3 months. X-ray showed that 65 cases had vertebral bone fusion, 39 had fiber fusion. All the patients were clinically healed and had no recurrence of tuberculosis at the follow-up for 2 years, and X-ray showed that all patients had vertebral bone fusion. Conclusion  Carefully treating the patients with complex and compound spinal tuberculosis, exactly handling the surgical opportunity and surgical technique on the basis of antituberculosis chemotherapy, we can achieve satisfactory results with classic debridement surgery.

Feasibility study of the Liquid-based Peng’s interlayer vessel technique on diagnosis of tuberculosis in primary level laboratory

SONG Hong-huan, JI Ming, TANG Guo-feng, ZHU Wei, ZHU Cheng-yu, CHEN Cheng，XU Wei-guo, LU Wei

Chinese Journal of Antituberculosis. 2012, 34(7):  441-444. 

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Objective  To evaluate the feasibility and security of Liquid-based Peng’s interlayer vessel technique used for diagnosis of tuberculosis in primary level laboratory.  Methods  In two primary level laboratories of Taixing and Jiangyin Center for Disease Control and Prevention, 754 sputum samples from 385 suspect TB patients were tested simultaneously by two methods: sputum smear and acid-fast staining microscopy examination (direct AFB) and Liquid-based Peng’s interlayer vessel technique. According to clinical diagnosis, the positive rates of the two methods for the sputum samples with different traits or collected at different times were compared by Fisher probabilities in 2×2 table. 12 bacterium-positive sputa were treated with different proportion of digestion liquid, different digestion time, and heating or not. Then the digestive sputa were inoculated at Roche medium at  37℃. The security of the liquid-based method was evaluated by preparing the growth of mycobacteria.  Results  The positive rates of Liquid-based Peng’s interlayer vessel technique and the direct AFB were 23.5% (177/754) and 16.8% (127/754) respectively, which had significant difference (χ²=48.02, P=0.000). The positive rates of these two methods in purulent sputa were 47.5% (94/198) and 40.9% (81/198), which had significant differences (P<0.001). The positive rates for the non-purulent sputa (including bloody sputa, mucus sputa and saliva sputa) were 13.5% (26/193) and 4.7% (9/193) (P<0.001). The sputum samples treated with the digestive liquid were not found the growth of Mycobacteria after the culture for 8 weeks.  Conclusion  Liquid-based Peng’s interlayer vessel technique can improve the positive detection rate in suspect TB patients. It is simple, safe, and suitable for the application at primary laboratory for the diagnosis of tuberculosis and convenient for quality control.

Correlation between MBL-A/B gene with the susceptibility of tuberculosis in Xinjiang Uighur population

ZHOU Jiang，ZHANG Wan-jiang

Chinese Journal of Antituberculosis. 2012, 34(7):  445-451. 

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Objective  To study the relationship between the polymorphism of the MBL gene and the susceptibility of tuberculosis (TB) in Xinjiang Uighur population.  Methods  The polymorphism of the MBL-A/B gene was analyzed by PCR-sequence specific primer (SSP) in 226 patients with pulmonary TB and 231 healthy controls who had a history of exposure to M. tuberculosis in Xinjiang Uighur population. The relationship between MBLgenotypes and the susceptibility of pulmonary TB was studied in accordance to the case-control study, and cases were grouped according to the genotypes.  Results  In the case group, AA, AB and BB genotypes at MBL gene A/B site were 106 cases (46.90%), 106 cases (46.90%) and 14 cases (6.20%), respectively. In the control group, AA, AB and BB genotypes were 146 cases (63.20%), 80 cases (34.63%) and 5 cases (2.17%), respectively. The frequency of BB genotypes at MBL gene A/B site in the case group was significantly higher than that in the healthy control group (6.20% vs 2.17%)(χ²=5.224，P_c＜0.05).  Conclusion  AA genotype of MBL gene may be protective factor for TB, while BB genotype may be risk factor for TB in Xinjiang Uighur population.

In vitro minimum inhibitory concentration of Mycobacterium tuberculosis isoniazid-resistant gene mutants

ZHANG Hai-qing,LIU Cheng-yong, SUN Qing, ZHANG Rui-mei, HUANG Hai-bin, HOU Yuan-pei,CHENG Song

Chinese Journal of Antituberculosis. 2012, 34(7):  452-454. 

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Objective  To study katG, inhA and ahpC mutations of Mycobacterium tuberculosis isoniazid-resistant clinical isolates, and their effect on minimum inhibitory concentration (MIC) in vitro.  Methods  Isoniazid-resistant genes of 160 Mycobacterium tuberculosis isoniazid-resistant strains and 40 isoniazid-sensitive strains were detected by gene chip. All samples with different gene mutations were determined and compared MICs of isoniazid.  Results  Of isoniazid-resistant strains, 68.8% （110/160） had katG 315 mutations, the MICs of katG 315 gene mutants （20.20±19.46 μg/ml）was significantly higher than inhA gene mutants （0.73±0.82 μg/ml, t′=10.9171, t′_α=1.9807,P＜0.05）, and significantly lower than katG 315 plus inhA gene mutants （69.33±32.45 μg/ml, t′= 7.164, t′_α= 2.0627, P＜0.05）.  Conclusion  Isoniazid-resistant strains had different gene mutation patterns, different genotypes had different levels of isoniazid resistance, which can provide a reference to the clinical.

Effect evaluation of the 5^th round of the Global Fund project on TB-HIV co-infection control in Xixiangtang district, Nanning city

ZHAO Ya-ling，LIN Xin-qin，QIN Xun，LIU Shao-ming

Chinese Journal of Antituberculosis. 2012, 34(7):  455-458. 

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Objective  To evaluate the effect of the 5^th round of the Global Fund project on TB-HIV co-infection control in Xixiangtang District, Nanning city.  Methods  We made a descriptive analysis on the quarterly report by the Fourth Peoples’ Hospital of Nanning city reported from October 2006 to September 2010, and carried out HIV antibody test for 3821 TB cases attending the hospital and TB screening for 4085 HIV/AIDS cases being followed-up.  Results  HIV antibody test proportion of TB cases was 72.5% (3821/5272); among them, 70 HIV positive cases were detected with the HIV-positive detection rate of 1.8% (70/3821). TB screening proportion of HIV/AIDS cases being followed-up was 92.2% (4085/4431); among them, 686 case were found to be TB cases with the TB detection rate of 16.8% (686/4085). From October 2006 to September 2009, there were 265 TB-HIV co-infection cases reported in total, of whom 7.6% （20/265） were cured, 44.5% （118/265） completed the treatment course, 24.5% (65/265) died and 23.4% (62/265) lost to follow-up.  Conclusion  We should strengthen the treatment management of co-infection patients and health education, and continuously improve the treatment success rate and reduce the loss rate.

Application of the grey model of metabolism for prediction of the pulmonary tuberculosis incidence in Guangdong province

JIANG Li, ZHONG Qiu, ZHOU Lin, LI Jian-wei, LIAN Yong-e

Chinese Journal of Antituberculosis. 2012, 34(7):  459-462. 

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Objective  To utilize the metabolizing grey model to predict the trend of TB incidence and provide scientific evidence for formulating the related measures of prevention and control.  Methods  According to the incidence of pulmonary TB in the Guangdong province from 2001—2011, we established the conventional grey model ［gray model (1,1), referred to as GM (1,1)］ and the grey model of metabolism ［referred to as the metabolic GM (1,1) model］. We compared the precision and accuracy of model predictions by the results of short series and long series, then chose the best model for extrapolation forecast. The prediction accuracy can be showed by posterior error ratio C and small probability of error P. The smaller class the best accuracy, fist class indicate the best and four failed.  Results  The error of the grey model of metabolism is less in comparison with the corresponding dimensions of conventional grey model. The error is less in the shorter series prediction. The result of accuracy test showed that the 5-dimensional grey model of metabolism is in the first class. The model of posterior error ratio and small probability of error was 0.14 and 1 respectively.  Conclusion  The metabolic GM (1,1) is the ideal model to deal with such data, the five-dimensional metabolic GM (1,1) model has certain advantages to forecast the TB incidence in Guangdong province.

Annual report on clinical diagnosis and treatment progress of tuberculosis (2011)（Part 2 clinical treatment）

Chinese Antituberculosis Association of Clinic Society

Chinese Journal of Antituberculosis. 2012, 34(7):  463-471. 

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