Serological diagnosis of pulmonary tuberculosis and nontuberculous pulmonary mycobacteriosis]

OBJECTIVE: The present study was undertaken to evaluate the utility of the serodiagnosis of pulmonary tuberculosis and nontuberculous pulmonary mycobacteriosis by ELISA using a Pathozyme-Myco kit (Myco kit) and a Pathozyme-TB complex kit (TB kit) (OMEGA Diagnostics Ltd.). STUDY POPULATION: The subjects comprised 256 healthy volunteers (HV, healthy hospital employees), 66 patients with sputum-positive active pulmonary tuberculosis (apTB), 14 patients with healed pulmonary tuberculosis (hpTB), 24 patients with nontuberculous pulmonary mycobacteriosis (NTM) and 32 patients with pulmonary diseases other than mycobacteriosis. RESULTS: 1) The serum IgG antibody titers determined with the Myco kit were significantly higher in the apTB group (p < 0.01), the hpTB group (p < 0.01), and the NTM group (p < 0.01) than those in the HV and the other pulmonary disease group. At a cut-off value of the mean + 2SD of the values obtained in the HV, the positive rate was 47.0% in patients with apTB, 50.0% in those with NTM, 21.4% in those with hpTB, 3.1% in those with other pulmonary diseases, and 1.6% in the HV. Analysis of ROC curves showed that the HV and the pulmonary mycobacteriosis group (apTB and NTM) were best distinguished by a cut-off value of -0.280 OD (log), with the sensitivity and the specificity being 83.3% and 78.5%, respectively. It was impossible to distinguish apTB from NTM. 2) The serum IgG antibody titers determined with the TB kit were significantly higher in the apTB group than those in the HV (p < 0.01), the NTM group (p < 0.05) and the other pulmonary disease group (p < 0.01). No significant difference was observed between the HV and the patients with NTM or those with other pulmonary diseases. Although the positive rate of the test was low in the apTB group (42.4%), there was a significant difference between apTB and NTM (12.5%) (p < 0.05), suggesting that apTB could be distinguished from NTM. 3) Since the serum antibody titers determined by the Myco kit showed no significant difference between apTB and NTM, and there was also no difference in the positivity between the two diseases, we performed serologic examination using the Myco kit to detect both diseases as pulmonary mycobacteriosis. After diagnosing pulmonary mycobacteriosis by the Myco kit, we then used the TB kit to separate apTB from NTM. In this case, the sensitivity and specificity of the test were 55.6% and 85.7%, respectively. Better methods should be developed to distinguish apTB from nontuberculous mycobacteriosis.     

Diagnostic role of fiberoptic bronchoscopy for mycobacterial diseases--serological diagnosis]

Detection of IgG antibodies against purified cord factor (trehalose-6, 6'-dimycolate) prepared from Mycobacterium tuberculosis H37Rv was carried out by the method of enzyme-linked immunosorbent assay (ELISA) and its diagnostic usefulness was also evaluated in this study. Sera from 65 patients with active pulmonary tuberculosis, 58 patients with inactive pulmonary tuberculosis, 36 patients with diseases other than tuberculosis and 66 healthy adults were examined. Patients with active pulmonary tuberculosis showed significantly higher titers of IgG antibodies against cord factor than other groups (p < 0.001). Patients with inactive pulmonary tuberculosis also showed significantly higher titers of IgG antibodies against cord factor than patients with diseases other than tuberculosis and healthy adults (p < 0.001). An antibody titers of greater than 0.29 were established as a positive ELISA test. For patients with active pulmonary tuberculosis, the ELISA had a sensitivity of 85% and a specificity of 96%. From these results, it is concluded that the detection of IgG antibodies against cord factor is useful for the serodiagnosis of active or inactive pulmonary tuberculosis.     

Humoral immune response against 38-kDa and 16-kDa mycobacterial antigens in tuberculosis.

Several ELISA tests based on mycobacterial antigens have been used for the rapid diagnosis of tuberculosis (TB), although demonstration of Mycobacterium tuberculosis in a smear or culture is the most reliable method. In the present study, the diagnostic value of 16-kDa and 38-kDa mycobacterial antigens was investigated in patients who were diagnosed with tuberculosis by clinical and/or bacteriological findings in Turkey. The PATHOZYME-TB Complex Plus commercial ELISA kit was used for measuring immunoglobulin G against 38-kDa and 16-kDa recombinant antigens. Humoral immune response was analysed in a group of 179 TB patients (143 smear-positive, 19 smear-negative, eight lymphadenitis and nine pleuritis), 15 inactive TB cases and in control groups consisting of 40 healthy volunteers and 20 subjects with pulmonary diseases other than TB. The sensitivity, specifity, positive predictive value and negative predictive value of the test were determined at 52.5%, 93.3%, 95.9% and 39.7%, respectively in TB cases. Antibodies were detected at above cut-off level in three (20%) out of 15 subjects with inactive TB. In conclusion, the ELISA test has a very good specifity and an acceptable sensitivity and positive predictive value. It is thought that it could be used in combination with other methods to increase diagnostic accuracy, especially for culture-negative tuberculosis cases, which are difficult to diagnose.     

Adenosine deaminase (ADA) in peritoneal tuberculosis: diagnostic value in ascitic fluid and serum

Simultaneous determination of ascitic fluid and serum adenosine deaminase (ADA) activity was evaluated as a diagnostic aid in peritoneal tuberculosis. The ascites was due to peritoneal tuberculosis (group 1), cirrhosis of the liver (group 2), cirrhosis of the liver with spontaneous bacterial peritonitis (group 3), peritoneal malignancy (group 4), Budd-Chiari Syndrome (group 5) and miscellaneous conditions (group 6). Serum from patients of pulmonary tuberculosis and healthy volunteers was analysed for enzyme activity. In patients with peritoneal tuberculosis the ascitic fluid and serum ADA activity was significantly higher than for the other groups (P less than 0.001). Levels above 36 u/l in ascitic fluid and above 54 u/l in the serum suggest tuberculosis. The ascitic fluid/serum ADA ratio was also higher in patients with peritoneal tuberculosis than with other causes of ascites (P less than 0.01). A ratio of more than 0.984 was suggestive of tuberculosis.     

结核分枝杆菌特异性蛋白抗体检测在结核病诊断中的价值

目的研究结核分枝杆菌特异性蛋白（TB-SA）抗体检测在结核病诊断中的价值。方法对829例结核病患者（男471例，女358例）、278例非结核性肺部疾病患者（男172例，女106例）和125例健康志愿者（男51例，女74例）同时进行结核菌素纯蛋白衍生物（PPD）试验，结核分枝杆菌TB-SA抗体检测，并对全部肺结核和非结核性肺部疾病患者进行痰抗酸杆菌涂片和培养。采用EPl2000统计软件进行统计学分析，组间比较采用t检验，计数资料采用X^2检验。结果结核分枝杆菌TB-SA抗体检测诊断菌阳肺结核的敏感度为75．1％（272／362）；诊断菌阴肺结核的敏感度为68．9％（226／328）；诊断肺外结核病的敏感度为71，2％（99／139）；诊断结核病的总体敏感度为72．0％（597／829），特异度为82．1％（331／403）。TB-SA血清抗体检测值并不与PPD值呈线性关系，即结核分枝杆菌TB-SA抗体检测诊断结核病不受卡介苗接种反应的影响。结论结核分枝杆菌TB-SA抗体检测诊断结核病有较好的敏感度和特异度，是辅助诊断和鉴别诊断结核病的有效手段。     

Sero-reactivity of two different antigenic protein fractions of M.tb H37Ra excretory-secretory antigen in pulmonary and extra-pulmonary tuberculosis

Excretory-secretory proteins of Mycobacterium tuberculosis H37Ra, have been of diagnostic interest in pulmonary (PTB) and extrapulmonary tuberculosis (EPTB). Two different excretory-secretory antigenic proteins of M.tbH37Ra viz., EST-DE1 (a 6% TCA soluble and DEAE anion exchange purified antigen) and ESAS-7 (50% ammonium sulphate solubilized and SDS-PAGE fractionated antigen) were studied in stick-indirect penicillinase ELISA for detecting tuberculous IgG antibodies in serum samples of pulmonary as well as extrapulmonary tuberculosis (tuberculous lymphadenopathy (TBLN), tuberculous meningitis (TBM), bone & joint tuberculosis (B&J TB), abdominal tuberculosis (Abd. TB) patients. The ESAS-7 antigen has shown comparatively better seroreactivity (90%) than that of EST-DE1 antigen in pulmonary tuberculosis cases. The overall specificity of 93.2% using ESAS-7 antigen was also found better compared to 86.4% obtained using EST-DE1 antigen. Further, in extra pulmonary tuberculosis group, using ESAS-7 antigen 84% (21/25) of histopathologically confirmed TBLN cases and 90% (9/10) clinically diagnosed and ATT responded TBM cases showed positive reaction for tuberculous IgG antibody. The per cent positivity using EST-DE1 antigen was however comparatively low in TBLN and TBM cases, (76% and 80% respectively). In histopathologically proven bone and joint tuberculosis and abdominal tuberculosis cases EST-DE1 antigen showed better sensitivity of 75% and 83.3% respectively in IgG antibody detection compared to that of ESAS-7 antigen (50% and 66% respectively). From the present study, it can be envisaged that ESAS-7 antigenic fraction has a good potential in the diagnosis of pulmonary and certain extra-pulmonary tuberculosis infection (TBLN & TBM) whereas EST-DE1 was found to be better in detecting specific antibodies in bone & joint and abdominal tuberculosis.     

ELISPOT检测在老年肺结核诊断中的应用价值

目的评价结核分枝杆菌特异性IFN-γ酶联免疫斑点(Elispot)检测技术对老年人肺结核的诊断价值。方法对46例老年肺结核、85例中青年肺结核、22例老年非肺结核、97例健康对照,进行Elispot检测并分析该技术诊断老年肺结核的可靠性。结果老年和中青年肺结核Elispot检测的敏感度分别为76.19%和87.5%,特异度分别为84.47%和85.29%。中青年和老年肺结核组Elispot检测阳性率显著高于其它两组(P<0.05)。结论 Elispot检测技术在诊断老年肺结核中显示出较好的敏感性和特异性,阳性率高,可用作老年肺结核的辅助诊断。     

Outcome following emergency surgery for refractory severe ulcerative colitis in a tertiary care centre in India

Background

The accurate diagnosis of abdominal tuberculosis usually takes a long time and requires a high index of suspicion in clinic practice. Eighty-eight immune-competent patients with abdominal tuberculosis were grouped according to symptoms at presentation and followed prospectively in order to investigate the effect of symptomatic presentation on clinical diagnosis and prognosis.

Methods

Based upon the clinical presentation, the patients were divided into groups such as non-specific abdominal pain & less prominent in bowel habit, ascites, alteration in bowel habit, acute abdomen and others. Demographic, clinical and laboratory features, coexistence of pulmonary tuberculosis, diagnostic procedures, definitive diagnostic tests, need for surgical therapy, and response to treatment were assessed in each group.

Results

According to clinical presentation, five groups were constituted as non-specific abdominal pain (n = 24), ascites (n = 24), bowel habit alteration (n = 22), acute abdomen (n = 9) and others (n = 9). Patients presenting with acute abdomen had significantly higher white blood cell counts (p = 0.002) and abnormalities in abdominal plain radiographs (p = 0.014). Patients presenting with alteration in bowel habit were younger (p = 0.048). The frequency of colonoscopic abnormalities (7.5%), and need for therapeutic surgery (12.5%) were lower in patients with ascites, (p = 0.04) and (p = 0.001), respectively. There was no difference in gender, disease duration, diagnostic modalities, response to treatment, period to initial response, and mortality between groups (p > 0.05). Gastrointestinal tract alone was the most frequently involved part (38.5%), and this was associated with acid-fast bacteria in the sputum (p = 0.003).

Conclusion

Gastrointestinal tract involvement is frequent in patients with active pulmonary tuberculosis. Although different clinical presentations of patients with abdominal tuberculosis determine diagnostic work up and need for therapeutic surgery, evidence based diagnosis and consequences of the disease does not change.     

sICAM-1 as a serum marker in the diagnosis and follow-up of treatment of pulmonary tuberculosis.

OBJECTIVE: To detect the value of sICAM-1 in the diagnosis and follow-up of treatment of tuberculosis. DESIGN: sICAM-1 levels were evaluated before and after treatment in 30 patients with pulmonary tuberculosis, only before treatment in five patients with pneumonia, five with lung cancer, and five with bronchiectasis, and in 10 healthy volunteers. RESULTS : sICAM-1 levels were as follows: 436.2 +/- 194.4 ng/ml in patients with pulmonary tuberculosis, 274 +/- 32.1 ng/ml in lung cancer patients, 268 +/- 41.9 ng/ml in bronchiectasis patients, 199.6 +/- 43.1 ng/ml in pneumonia patients, and 146.5 +/- 20.2 ng/ml in healthy individuals. sICAM-1 levels of tuberculosis cases before treatment were higher than in both the healthy group and in all the other groups. The levels in the healthy group were lower than in all other groups. CONCLUSION: The cut-off point (298 ng/ml) obtained by adding the standard deviation to the mean sICAM-1 value of patients without tuberculosis had 83.3% sensitivity, 86.6% specificity and 84.4% accuracy in differentiating pulmonary tuberculosis from other pulmonary diseases. sICAM-1 can be used as an auxiliary marker in the diagnosis of pulmonary tuberculosis.     

结核分支杆菌重组38 000蛋白的免疫原性特征

目的了解结核分支杆菌重组３８０００蛋白（ｒＭＴ３８）诱发家兔产生抗体的能力及作为血清学诊断试剂的价值。方法将家兔分为６组：ＰＰＤ＋生理盐水（Ａ组）,ｒＭＴ３８＋生理盐水（Ｂ组）ｒＭＴ３８＋佐剂（Ｃ组）,１／２ｒＭＴ３８＋１ｒＭＴ１６＋生理盐水（Ｄ组）,１／２ｒＭＴ３８＋１／２ｒＭＴ１６＋佐剂（Ｅ组）,ｒＭＴ１６＋佐剂（Ｆ组））。用ＥＬＩＳＡ法测Ｃ、Ｅ两组血清是否与分支杆菌ＰＰＤ有效叉反应。双向扩散     

Detection of anti-cord factor antibodies in intestinal tuberculosis for its differential diagnosis from Crohn's disease and ulcerative colitis

We have developed a diagnostic method for pulmonary tuberculosis by detecting antibody to cord factor using enzyme-linked immunosorbent assay (ELISA). This study was to evaluate the usefulness of our method for a diagnosis of intestinal tuberculosis, and especially its ability to differentiate this disease from other inflammatory bowel diseases. Antibodies of the immunoglobulin G class against cord factor (trehalose-6,6'-dimycolate) from 27 patients with intestinal tuberculosis, 16 patients with Crohn's disease (CD), and 27 patients with ulcerative colitis (UC) were tested by ELISA with cord factor purified fromMycobacterium tuberculosis H37Rv as the antigen. Twenty-three of the 27 patients with intestinal tuberculosis (85%) showed elevated values distinct from healthy controls. None of the patients with CD showed an elevation of antibody titers. Of the 27 patients with UC, 26 (96%) did not show any anti-cord factor antibody elevation. We conclude that this method is simple and results are reproducible. The results of our study justify undertaking the detection of anti-cord factor antibodies to diagnose intestinal tuberculosis.     

重组结核分支杆菌38kD和16kD蛋白用于结核病血清学诊断的价值

目的　探讨国产重组结核分支杆菌蛋白38kD(rTPA38)和16kD(rTPA16)用于结核病的诊断价值。方法 以rTPA38和rTPA16为抗原,PPD为对照,用ELISA法检测血清中特异性抗结核抗体。结果 246例肺结核病组,rTPA38、rTPA16和PPD检测的灵敏度分别为66.3%、63.0%和72.3%。健康献血组、非结核呼吸疾病组和卡介苗接种阳转组血清同时用rTPA38、rTPA16和PPD检测,rTPA38特异性分别为97.6%、96.8%、86.0%。rTPA16特异性分别为94.7%、93.1%、75.0%。PPD特异性为93.4%、85.7%、67.9%。统计分析显示,rTPA38蛋白和PPD检测非结核呼吸疾病组,其阳性率有显著性差异 (P＜0.05)。两者检测卡介苗接种阳转组阳性率和血清抗体滴度有显著性差异 (P＜0.05)。rTPA38和rTPA16同时检测108例肺结核病组血清可提高11.1%的阳性率。结论 rTPA38蛋白抗原有较好的灵敏度和较高特异性,是ELISA的可靠抗原,与rTPA16联用可提高灵敏度,对结核病血清学诊断有较高参考价值。     

CA125检测在活动性结核判定及疗效评估中的价值

目的　探讨血清CA12 5测定在活动性肺结核判定及疗效评估中的价值。方法　应用化学发光法分别对3 0例活动性肺结核治疗前、治疗后 2个月、4个月、6个月以及 3 0例非活动性肺结核和 3 0例健康体检者血清CA12 5进行测定。并对结果进行分析。结果　活动性肺结核治疗前血清CA12 5水平显著高于非活动性肺结核及健康体检者 (P <0 0 1)。活动性肺结核治疗前CA12 5水平显著高于治疗后各期测定值 (P <0 0 1) ,且治疗后血清CA12 5水平呈递减趋势。活动性肺结核治疗后 6个月血清CA12 5水平与非活动性肺结核比较无明显差异 ( P >0 0 5 )。结论　血清CA12 5测定可作为判定活动性肺结核及评估疗效的可靠指标     

Value of enzyme immunoassay in the diagnosis and differential diagnosis of abacillary tuberculosis in children and teenagers

Positive enzyme immunoassay (EIA) was found in 59.6% of patients of tuberculosis. Positive responses were significantly more frequently detected in patients with pulmonary tuberculosis than in those with intrathoracic lymph nodal tuberculosis (ITLNT) (62.2 +/- 7.4 versus 38.9 +/- 11.8%, respectively; p < 0.05. There was no significance difference in the rate of positive responses in patients with ITLNT (38.9 +/- 11.8%) and healthy individuals inoculated with Mycobacterium tuberculosis (17.6 +/- 9.5%). Sixteen (61.5%) patients with pneumonia showed a positive response, which amounted to 38.5% specificity of the method. The findings suggest that EIA may not be used as a screening test for early diagnosis of tuberculosis in children and teenagers and for differential diagnosis of pulmonary tuberculosis and nonspecific pneumonia. EIA data may be assessed only in combination with other diagnostic criteria.     

Usefulness of measuring serum IGG antibodies against A60 mycobacterial antigen for diagnosis of tuberculosis

Measurement of antimycobacterial antibody may be used as potential diagnostic tool in tuberculosis. The aim of the study was to evaluate the diagnostic value of serum IgG level against A60 mycobacterial antigen measured by ELISA method. Material consisted of 144 persons divided into 5 groups (76 tuberculosis patients, 20 sarcoidosis patients, 17 lung cancer patients, 8 patients with mycobacterial infections other than tuberculosis and 23 healthy controls). In the tuberculosis group there were 50 culture positive cases and 26 culture negative ones, 43 new cases and 32 chronic cases. Positive results were obtained in 51% of tuberculosis patients. Sensitivity increased to 62% in culture positive group and 63% in chronic cases. Specificity of the test was 96%. The results indicate that Immunozyme Mycobacterium test is a valuable tool in tuberculosis diagnosis.     

抗结核抗体检测方法及临床意义

目的 探讨抗结核抗体检测技术在临床诊断中的应用价值.方法 对1823例确诊肺结核患者、580例非结核病的其他呼吸系统疾病患者及500名健康体检者进行抗酸染色法、酶联免疫吸附试验(ELISA)法、金标法及蛋白芯片法检测,并进行相关统计分析.结果 ELISA法、金标法及蛋白芯片法对肺结核患者的检出敏感性分别为59.1%、62.7%和71.2%,均显著高于抗酸染色(18.4%).蛋白芯片法对抗酸染色阴性肺结核患者的抗结核抗体检出率为67.9%.结论 3种结核抗体检测技术均可用于肺结核的快速辅助诊断.相对于ELISA法及金标法而言,蛋白芯片法敏感性和特异性更高,对痰菌阴性肺结核患者的辅助诊断更具积极意义.     

The value of Ca 125 in the evaluation of tuberculosis activity.

The aim of the present study was to investigate the value of Ca 125, a tumour marker, in evaluation of pulmonary tuberculosis activity. This study included 96 subjects who were divided into three groups. Group 1 consisted of 40 patients with active pulmonary tuberculosis. Group 2 included 20 patients with inactive pulmonary tuberculosis. There were 36 healthy subjects in group 3. While measurement of serum Ca 125 level was performed only once in groups 2 and 3, Ca 125 levels were measured five times in group 1. The measurements were performed before the treatment, at the second, fourth and sixth months and the third year following the end of the treatment. Mean +/- SD serum Ca 125 concentrations were 109.7 +/- 86.9 U ml(-1) in group 1, 14.5 +/- 7.8 U ml(-1) in group 2 and 10.5 +/- 7.3 U ml(-1) in group 3. Serum Ca 125 levels were significantly higher in group 1 than in the other groups (P < 0.0001), but there was no significant statistical difference between the values of groups 2 and 3 (P > 0.05). Ca 125 levels in group 1 showed a significant decrease after treatment (P < 0.0001). For estimation of the activity of tuberculosis, the sensitivity and specificity of Ca 125 were found 97.5% and 100%, respectively at a 31 U ml(-1) cut-off point. Our results suggest that Ca 125 is beneficial in the determinaton of tuberculosis activity and in differentiation between active and inactive pulmonary tuberculosis.     

慢性阻塞性肺疾病急性加重期合并肺结核患者外周血清中IL-6、TNF-α和IFN-γ水平变化及临床意义

目的 探究慢性阻塞性肺疾病急性加重期(AECOPD)合并肺结核患者外周血清中IL-6、肿瘤坏死因子α(TNF-α)和干扰素γ(IFN-γ)水平变化及临床意义.方法 将研究对象分为AECOPD合并肺结核组、AECOPD组、肺结核组和健康对照组,根据肺结核患者影像学检查结果将肺结核患者分为单侧病变组、双侧病变组、无空洞组和有空洞组,根据AECOPD合并肺结核患者治疗后的情况分为好转组和恶化组.采用ELISA法检测各组患者血清中的IL-6、TNF-α和IFN-γ水平.结果 AECOPD合并肺结核组血清中的IL-6和TNF-α水平显著高于AECOPD组和肺结核组(P＜0.05);单侧病变组患者血清中的IL-6、TNF-α和IFN-γ的水平显著低于双侧病变组(P＜0.05);无空洞组患者血清中的IL-6、TNF-α和IFN-γ的水平显著低于有空洞组(P＜0.05);病情好转组治疗后IL-6、TNF-α和IFN-γ水平显著低于治疗前(P＜0.05).结论 IL-6、TNF-α和IFN-γ在AECOPD合并肺结核的发病机制中起着一定的作用,其外周血清中水平的动态变化在一定程度上可以反映AECOPD合并肺结核病情的发展与转归.     

Diagnostic evaluation of the determination of antibodies to purified protein derivative in tuberculosis]

The results with ELISA method showed the levels of the antibody to PPD in exudates of TB group were significantly higher than those of non-TB group (P less than 0.001). When OD value greater than 0.2 and greater than 0.35 were chosen as the positive criteria, for diagnosing tuberculous meningitis and other tuberculous, the sensitivity would be 79.3% and 86.6%, respectively; the specificity would be 93.3% and 86.0%, respectively. Meanwhile tuberculosis could be suggested when the ratio of the antibody level in exudate to that in serum is greater than 1. The authors think that the detection of the antibody to PPD in exudates might be a supplemental diagnostic tool for tuberculosis.