Hysteroscopic observations in 348 consecutive cases of female genital tuberculosis: A prospective study

Study objectiveTo evaluate the hysteroscopic findings in female genital tuberculosis.DesignIt was a prospective study of hysteroscopic findings performed on 348 cases of female genital tuberculosis (FGTB).SettingIt was a prospective cross-sectional study in a tertiary referral centre.PatientsA total of 348 patients with infertility with FGTB on various tests.InterventionA total of 348 patients of infertility found to have FGTB on various investigations were enrolled in the study. A detailed history was taken. Clinical examination, endometrial sampling and diagnostic laparoscopy were performed was also performed in selected cases. All patients underwent hysteroscopy as part of evaluation for infertility and tuberculosis (TB) findings.Measurements and main resultsThe mean age, parity, body mass index and duration of infertility was 28.2 years, 0.31,23.1kg/m² and 3.44 years respectively. Infertility was primary in 81.03% and secondary in 18.96% cases. Diagnosis of FGTB was made by endometrial aspirate findings of positive AFB on microscopy (4.02%), positive culture (4.88%), positive PCR (83.90%), epithelioid granuloma (14.65%), positive AFB on microscopy or culture of peritoneal cytology (1.14%) or epithelioid granuloma on peritoneal biopsy (1.72%), definitive findings of TB on laparoscopy (41.95%) or probable findings of TB on laparoscopy (58.05%). Various hysteroscopic findings observed were normal findings (28.16%), pale endometrial cavity (54.31%), features of active TB (7.47%), features of chronic TB (19.54%), features of TB sequelae like obstructed ostia (both ostia in 13.79%, one ostia 14.94%, periostial fibrosis; (bilateral 4.59%, unilateral 5.17%), endometrial glands atrophy (12.35%), small shrunken cavity (6.32%), distorted cavity (5.17%), various grades of intrauterine adhesions (29.88%). Hysteroscopy in FGTB was associated with increased difficulties and complications like failed procedures, difficult visualisation, false passage and uterine perforation.ConclusionHysteroscopy is useful modality to detect endometrial TB but is associated with increased difficulty and complications.     

Computed tomographic findings in female genital tuberculosis tubo-ovarian masses

Female genital tuberculosis (FGTB) is a common cause of infertility in developing countries. It can manifest as menstrual disturbances, infertility and pelvic masses.ObjectiveTo evaluate the role of computed tomography in diagnosis of female genital tuberculosis with tubo-ovarian (adnexal) masses.MethodsIt was a prospective study over a four year period (July 2015 to August 2019) in a tertiary referral centre over 33 patients presenting with tuberculosis and tubo ovarian masses only. 75 total cases of FGTB diagnosed on composite reference standard (evaluation of AFB bacilli in microscopy or culture or endometrial biopsy, gene expert, epitheloid granulomas on endometrial biopsy or definitive or possible findings of FGTB on laparoscopy). Detailed history taken, clinical examination, baseline investigations and endometrial biopsy were done in all cases. Computed tomography was performed in women presenting with infertility, tubo ovarian masses on clinical examination and laboratory investigations. A total of 33 cases were evaluated.ResultsMean age, body mass index, parity and history of TB contact were 27.5 ± 4.2 year, 22.7 ± 3.6 kg/m², 0.27 ± 0.13 and 44.4% respectively. Infertility was primary in 72.72% and secondary in 27.23%. Case wise mean duration being 5.8 years, menstrual dysfunction was seen in 45.45% cases. Abdominal discomfort with pain and lump were seen in all 33 (100%) cases. Abdominal lumps were felt in 4 (12.12%) cases while adnexal mass was seen in all 33 (100%) cases being unilateral in 18 (54.54%) and bilateral in 15 (45.45%). Mean ESR was 33.4mm in first hour while mean leucocyte count was 6128 ± 2854 per cubic mm. Infectious mantoux test (>10mm) was seen in 14 (42.82%) cases while abnormal X ray chest was seen in 9 (27.27%) cases.Diagnosis of FGTB was made by positive AFB n microscopy or culture of endometrial biopsy in 5 (15.15%) cases, positive gene expert in 6 (18.18%) cases, positive polymerase chain reaction in 32 (96.96%) cases, epitheloid granulomas on histopathology of endometrial biopsy in 7 (21.21%) cases, definitive findings of tuberculosis in 15 (45.45%) cases and a possible findings of tuberculosis inn 18 (54.54%) cases. Various CT findings were pelvic mass (100%), unilateral pelvic mass in 18 (54.54%), bilateral pelvic mass in 15 (45.45%), cystic mass (24.2%), solid mass (21.2%), mixed mass (54.54%), mass showing multilocular caseous necrotic enhancements (12.12%), ascites (42.4%), thickening and enhancement of peritoneum in 14 (42.42%), nodules in 24.2%, smooth in 18.8%, pelvic adhesion in 6 (18.18%), lymphadenopathy in 8 (24.3%) with calcifications (9.09%) and central necrosis (52.5%). Other CT findings were thickening and enhancement of bowel wall (12.12%), hepatic TB (3.03%), splenic TB (3.03%), omental thickening (9.09%) and omental calcification (3.03%) cases.ConclusionComputed tomography appears to be a useful diagnostic modality in diagnosis of tuberculosis tubo ovarian masses and may help avoid unnecessary surgery.     

An evaluation of Composite Reference Standard (CRS) for diagnosis of Female Genital Tuberculosis

BackgroundFemale genital tuberculosis (FGTB) is a common cause of infertility in developing countries. Its diagnosis is difficult due to its paucibacillary nature, with no single test having high sensitivity and specificity. This study is to share the experience of using Composite Reference Standard (CRS) for the diagnosis of FGTB.MethodsThis is a prospective study conducted between September 2017 to June 2019, over 100 infertile females found to have FGTB on composite reference standard which consisted of acid-fast bacilli on microscopy or culture, histopathological evidence of epithelioid granuloma, positive gene Xpert on endometrial sample or definite or probable finding of FGTB on laparoscopy.ResultsA total of 100 infertile women (78% primary, 22% secondary) found to have FGTB on CRS were enrolled in this study. Mean age, body mass index, parity and duration of infertility were 28.2 years, 23.17 kg/m², 0.24 ± 0.12 and 2.41 years respectively. Various symptoms were scanty menses (16%), irregular cycle (7%), dysmenorrhea (11%), pelvic pain (11%). Various signs were vaginal discharge (65%), adnexal mass (6%), tubo-ovarian mass on ultrasound (15%), abnormal hysterosalpingography findings (57.14%), positive polymerase chain reaction test (65%) and abnormal hysteroscopy (82.2%). The positive findings on CRS were positive AFB on microscopy or culture (3%), positive gene Xpert (28%) (done in some cases), epithelioid granuloma on histopathology (13%), definite findings on laparoscopy like tubercles, caseous nodules and beaded tubes in (57.19%) patients while probable findings of FGTB like straw colored fluid in POD, extensive dense pelvic, peri-tubal, peri-ovarian adhesions; hydrosalpinx; tubo-ovarian mass; thick fibrosed tubes; mid tubal block; peri hepatic adhesions (Fitz Hugh Curtis Syndrome); hyperemia of tubes/blue uterus on chromotubation were seen in (48.8%) patients. All patients found to be positive on CRS were given 6 months of anti-tubercular therapy.ConclusionThis study demonstrates the high reliability of use of composite reference standard for diagnosis of FGTB.     

Cartridge based nucleic acid amplification test is superior in diagnosing lymphnode tuberculosis

BackgroundThe role of Cartridge based Nucleic Acid Amplification test (CBNAAT) in the diagnosis of lymphnode TB which helps in reducing the mortality and morbidity by early identification and initiating treatment at the earliest. Also helps in identify the drug resistance among tubercular lymphnodes cases.Patients and methodsA prospective clinical study was performed in 101 suspected lymph node tuberculosis patients. The results of FNAC and/or excision biopsy of lymphnode samples obtained by CBNAAT were compared with direct smear microscopy for AFB bacilli, cytology and their combination considering AFB culture as gold standard.ResultsA total of 101 patients were evaluated of which 74 subjects (73.3%) were CBNAAT positive for TB. Among the CBNAAT positive cases, 57 were aged above 16 years, 38 were females, equal number (37) had single and multiple lymphnodes, 46 had less than 1 cm size lymphnodes, 69 had lymphnode in neck region, 65 had chest X-ray normal. Among CBNAAT positive 74 subjects, 53 subjects (71.6%) were positive for AFB direct smear, 64 subjects (86.48%) were cytology consistent with TB and their combination were positive for TB in 71 subjects (95.94%) and 71 subjects (95.94%) were positive by AFB culture and 3 cases (0.04%) showed Rifampicin resistance.ConclusionCBNAAT is a rapid diagnostic tool having sensitivity of 93.42% with specificity of 86.96% and positive predictive value of 95.95% and having comparable results with AFB culture and more sensitive than other investigation procedures. Thus it can be a rule in test for lymphnode TB.     

Laboratory diagnosis of pulmonary tuberculosis in TB and HIV endemic settings and the contribution of real time PCR for M. tuberculosis in bronchoalveolar lavage fluid

Background Tuberculosis (TB) in Africa is increasing because of the human immunodeficiency virus (HIV) epidemic, and in HIV/AIDS patients it presents atypically. Pulmonary tuberculosis (PTB) in Africa is mainly diagnosed clinically, by chest radiograph or by sputum smear for acid fast bacilli (AFB). Methods We evaluated in 120 HIV‐infected patients with chest infection the diagnostic accuracy of AFB smear of sputum and bronchoalveolar lavage (BAL) fluid, sputum Mycobacterium tuberculosis (MTB) culture, real‐time PCR and MycoDot^® serological test, using MTB culture of BAL fluid as gold standard. We correlated PCR cycle threshold values (C_T) to the culture results. Retrospectively, we evaluated the development of active TB in patients with positive PCR but negative culture. Results Culture of BAL fluid identified 28 patients with PTB. Fifty‐six patients could not produce adequate sputum. Sputum AFB smear and the serological test had sensitivities of 66.7% and 0%, respectively. PCR with C_T 40 was positive in 73 patients, 27 of whom were also TB culture positive (96.4% sensitivity and 52.3% specificity of PCR). PCR with C_T 32 had sensitivity of 85.7% and specificity of 90.9% to diagnose PTB in BAL. No patients with positive PCR but negative culture developed active TB during 18 months follow‐up. Conclusion In these HIV‐infected patients, AFB smear and serology had very low sensitivities. PCR of BAL with C_T value 32 had improved specificity to diagnose active PTB. A prospective follow‐up study is warranted in TB/HIV endemic settings, applying real time PCR to both sputum and BAL.     

Is the rise in Crohn's disease in India accompanied by a fall in intestinal tuberculosis? A single-center experience

IntroductionThe relationship between the incidence of intestinal tuberculosis (TB) and Crohn's disease (CD) is interesting, especially considering the striking similarity between the two conditions. Some studies from Asian populations suggested that the incidence of intestinal TB decreases when there is an increase in CD.AimTo compare the incidence trend between intestinal TB and CD over 15 years.MethodsMedical records of patients seen in the Division of Gastroenterology over 15 years (2005–2019) were reviewed. CD was diagnosed according to the Copenhagen criteria. Intestinal TB was diagnosed in the appropriate clinical situation if any one or more of the following was present: (1) positive TB MGIT culture; (2) positive Gene Xpert for TB; (3) suggestive histologic findings, with positive tissue acid-fast bacillus (AFB) on smear or with sustained response to anti-TB therapy. The incidence time trend of patients with CD and intestinal TB diagnosis was then studied year-wise.Results632 medical case records were accessed; 60 patients were excluded due to inadequate data or not fulfilling diagnostic criteria. The 572 patients included 224 with intestinal TB (median age 37 years, IQR 22; 125 [56%] females) and 348 with CD (median age 40 years, IQR 25; 159 [46%] females [p < 0.02 as compared to TB]). Thus, more patients with CD were seen during the study period, but there was no correlation between the incidence of the two conditions (r = 0.318; p = 0.25).ConclusionIn Indian patients in a single private-sector center, there was no inverse correlation between the incidence of intestinal TB and CD over 15 years.     

Regulatory T cells are expanded in blood and disease sites in patients with tuberculosis

RATIONALE: T-cell responses during tuberculosis (TB) help contain Mycobacterium tuberculosis in vivo but also cause collateral damage to host tissues. Immune regulatory mechanisms may limit this immunopathology, and suppressed cellular immune responses in patients with TB suggest the presence of regulatory activity. CD4+CD25(high) regulatory T cells mediate suppressed cellular immunity in several chronic infections but have not been described in TB. OBJECTIVE: To determine whether regulatory T cells are increased in patients with TB and whether they suppress cellular immune responses. METHODS: We compared the frequency of circulating regulatory T cells in 27 untreated patients with TB and 23 healthy control subjects using two specific markers: cell-surface CD25 expression and FoxP3 mRNA expression in peripheral blood mononuclear cells. MEASUREMENTS AND MAIN RESULTS: We detected a threefold increase in the frequency of CD4 + CD25(high) T cells (p < 0.001) and a 2.2-fold increase in FoxP3 expression (p = 0.006) in patients with TB, and there was a positive correlation between these markers (r = 0.58, p < 0.001). Increased expression of interleukin-10 and transforming growth factor-beta1 mRNA was also detected in patients with TB but did not correlate with regulatory T-cell markers. Ex vivo depletion of CD4 + CD25(high) cells from peripheral blood mononuclear cells resulted in increased numbers of M. tuberculosis antigen-specific IFN-gamma-producing T cells in seven of eight patients with TB (p = 0.005). Finally, FoxP3 expression was increased 2.3-fold in patients with extrapulmonary TB compared with patients with purely pulmonary TB (p = 0.01) and was amplified 2.6-fold at disease sites relative to blood (p = 0.043). CONCLUSIONS: Regulatory T cells are expanded in patients with TB and may contribute to suppression of Th1-type immune responses.     

Effector functions of heparin-binding hemagglutinin-specific CD8+ T lymphocytes in latent human tuberculosis

BACKGROUND: Most individuals infected with Mycobacterium tuberculosis do not develop tuberculosis (TB) and can be regarded as being protected by an appropriate immune response to the infection. The characterization of the immune responses of individuals with latent TB may thus be helpful in the definition of correlates of protection and the development of new vaccine strategies. The highly protective antigen heparin-binding hemagglutinin (HBHA) induces strong interferon (IFN)- gamma responses during latent, but not active, TB. Because of the recently recognized importance of CD8(+) T lymphocytes in anti-TB immunity, we characterized the CD8(+) T lymphocyte responses to HBHA in subjects with latent TB. RESULTS: HBHA-specific CD8(+) T lymphocytes expressed memory cell markers and synthesized HBHA-specific IFN- gamma . They also restricted mycobacterial growth and expressed cytotoxicity by a granule-dependent mechanism. This activity was associated with the intracellular expression of HBHA-induced perforin. Surprisingly, the perforin-producing CD8(+) T lymphocytes were distinct from the IFN- gamma -producing CD8(+) T lymphocytes. CONCLUSION: During latent TB, the HBHA-specific CD8(+) T lymphocyte population expresses all 3 effector functions associated with CD8(+) T lymphocyte-mediated protective immune mechanisms, which supports the notion that HBHA may be protective in humans and suggests that markers of HBHA-specific CD8(+) T lymphocyte responses may be useful in the monitoring of protection.     

Diagnosis of Crohn＇s disease in India where tuberculosis is widely prevalent

AIM： To define the parameters that positively predict diagnosis of Crohn＇s disease （CD） and differentiate it from gastrointestinal tuberculosis （GITB）.
METHODS： This prospective study over 3 years was carried out in the consecutive Indian patients with definite diagnosis of CD and equal numbers of patients with definite diagnosis of GITB. Demographic, clinical, laboratory, morphological and histological features were noted in all the patients. Serological tests such as p-ANCA, c-ANCA, IgA ASCA and IgG ASCA, were performed. Endoscopic biopsy and/or surgical tissue specimens were subjected to smear and culture for acid-fast bacilli （AFB） and tissue polymerase chain reaction for tuberculosis （TB PCR）. Diagnosis of CD and GITB was based on the standard criteria. Data were analyzed using univariate Chi-square test and multiple logistic regression （MLR）.
RESULTS： The study is comprised of 26 patients with CD （age 36.6 ± 8.6 year, male：female, 16：10） and 26 patients with GITB （age 37.2 ± 9.6 year, male：female, 15：11）. The following clinical variables between the two groups （CD vs TB） were significant in univariate analysis： duration of symptoms （58.1 ± 9.8 vs 7.2 ± 3.4 mo）, diarrhoea （69.2% vs 34.6%）, bleeding per rectum （30.7% vs 3.8%）, fever （23.1% vs 69.2%）, ascites （7.7% vs 34.6%） and extra-intestinal manifestations of inflammatory bowel disease （61.5% vs 23.1%）. Of these, all except ascites and extra-colonic manifestations were found statistically significant by MLR. Accuracy of predicting CD was 84.62% based on the fever, bleeding P/R, diarrhoea and duration of symptoms while it was 63.4% when histology was reported as inflammatory bowel disease and 42.3% when there was recurrence of disease after surgery. Accuracy of predicting GITB was 73.1% when there was co-existing pulmonary lesions and/or abdominal lymphadenopathy; 75% when tuberculosis was reported in histology; 63.4% when granuloma was found in histology; 82.6% when TB PCR was     

Importance of eosinophilic structures and its correlation with acid fast bacilli in fine needle aspiration smears of tubercular lymphadenitis

BackgroundThe hallmark of tuberculosis in cytological smears is the presence of epithelioid cell granulomas, necrosis and AFB. In cases, where AFB not detected, diagnosis of tuberculosis can be made by ancillary tests like PCR which requires infrastructure besides being expensive. In India, where majority of population is rural-based there is an imminent need of some morphological change in cytological smears which can point towards diagnosis of tuberculosis in absence of AFB. This study was done to assess the importance of eosinophilic structures (ES) and its correlation with presence of AFB.Material & methodsThis was a retrospective study over a period of one year. Lymph node aspirates reported as granulomatous lymphadenitis, tubercular lesion or suppurative lesion were included. All smears for each case, stained with May Grunwald Giemsa (MGG), Hematoxylin and Eosin (H&E) and ZN stain were retrieved and rescreened for the presence of eosinophilic structure, necrosis, granulomas and AFB.ResultsOur study included 256 cases. Most common age group was 21–30 years with female predominance. Cervical lymph nodes were most commonly involved. In relation to ES and AFB four cytological pictures were seen i.e ES + AFB+ (44.54%) ES + AFB- (5.46%) ES- AFB+ (14.45%) ES - AFB- (35.55%). Chi square test showed a high significant statistical association between ES and AFB(p = 0.00001).ConclusionIn developing countries with very high TB prevalence like India, where new diagnostic modalities are unaffordable by rural population, presence of ES should be included as a morphological parameter in routine cytology reporting of tubercular lesions.     

Drug resistance of Mycobacterium tuberculosis in selected urban and rural areas in Bangladesh

The magnitude of anti-tuberculosis drug resistance in Bangladesh is not precisely known. We studied the drug resistance patterns of Mycobacterium tuberculosis in an urban and a rural area of Bangladesh. A tuberculosis (TB) surveillance system has been set up in a population of 106,000 in rural Matlab and in a TB clinic in urban Dhaka. Trained field workers interviewed all persons > or =15 y at Matlab to detect suspected cases of tuberculosis (cough >21 d) and sputum samples were examined for acid-fast bacilli (AFB). The first 3 AFB positive patients daily from the urban clinic were included. AFB positive cases diagnosed between June 2001 and June 2003 from both settings were cultured and drug susceptibility tests were performed. Of 657 isolates, resistance to 1 or more drugs was observed in 48.4% of isolates. Resistance to streptomycin, isoniazid, ethambutol and rifampicin was observed in 45.2%, 14.2%, 7.9% and 6.4% of isolates, respectively. Multidrug resistance was observed in 5.5% of isolates. It was significantly higher among persons who previously had received tuberculosis treatment of > or =1 month (15.4% vs 3.0%, adjusted OR: 6.12, 95% CI: 3.03-12.34). The magnitude of anti-tuberculosis drug resistance in Bangladesh is high. Further evaluation is needed to explain the high proportion of streptomycin resistant M. tuberculosis. Appropriate measures to control and prevent drug resistant tuberculosis in Bangladesh to reduce mortality and transmission are warranted.     

结核感染T细胞斑点试验在不同患者群中的检测结果分析

目的 探讨影响结核感染T细胞斑点试验(T-SPOT.TB)检测结果的因素。方法收集2014年5月至2015年4月同济大学附属上海市肺科医院共1537例住院患者的临床资料。以《临床诊疗指南:结核病分册》的标准将患者分为临床诊断结核病组(1159例)和临床诊断非结核病组(378例);再经病原学诊断及随访排除不能做出最终诊断的39例患者后,将患者分为结核病组(1103例;包括经分枝杆菌培养及菌种鉴定确诊的229例和最终临床诊断的874例)和非结核病组(395例;包括既往有结核病史或肺部存在陈旧性病灶者94例,无病史者301例);1103例结核病组患者中,肺结核557例,结核性胸膜炎107例,淋巴结结核16例,骨关节结核51例,脑结核7例,多组织器官结核363例,其他肺外结核2例;395例非结核病组患者中,经分枝杆菌培养及菌种鉴定确诊的非结核分枝杆菌(NTM)感染患者93例。回顾性分析临床诊断结核病组和结核病组患者T-SPOT.TB检测结果的敏感度、特异度,结核病组与非结核病组患者T-SPOT.TB检测反应强度,不同结核病类型T-SPOT.TB检测阳性率,以及NTM感染患者与结核病患者的年龄差异。结果 (1)临床诊断结核病组T-SPOT.TB检测的敏感度为81.97% (950/1159)、特异度为53.44%(202/378);结核病组T-SPOT.TB检测的敏感度为83.77%(924/1103)、特异度为54.43%(215/395);结核病组中抗酸染色阳性患者T-SPOT.TB检测阳性率为90.73%(235/259);培养阳性患者T-SPOT.TB检测阳性率为92.58%(212/229)。(2)非结核病组有结核病史或肺部显示陈旧性结核病灶的患者与无病史者T-SPOT.TB检测的阳性率分别为69.15%(65/94)和38.21%(115/301),两组间差异有统计学意义(χ ²=27.65,P=0.000)。(3)结核病组与非结核病组T-SPOT.TB检测结果以斑点计数表示反应强度,结核病组对A、B抗原刺激出现阳性反应(斑点数≥1)的患者中,明显阳性及超强阳性(斑点数≥11)的比率分别为69.81%(652/934)和69.67%(627/900),明显高于非结核病组[分别为46.83%(96/205)和46.63%(83/178)];经曼-惠特尼秩和检验,差异有统计学意义(Z值分别为-14.20、-14.63,P值均<0.01)。(4)1103例结核病组患者中,T-SPOT.TB检测阳性率以淋巴结结核(87.50%,14/16)为最高,随后依次为多组织器官结核(87.33%,317/363)、肺结核(86.54%,482/557)、结核性胸膜炎(74.77%,80/107),脑结核(57.14%,4/7)和骨关节结核(50.98%,26/51)相对较低。(5)T-SPOT.TB检测阳性的NTM感染患者的平均年龄[(53.61±18.43)岁]较T-SPOT.TB检测阳性的确诊结核病患者[(44.98±18.88)岁]高,差异有统计学意义(t=-2.63,P=0.009)。结论 T-SPOT.TB检测结果受结核病诊断依据、痰菌量、既往结核病史或肺部陈旧性结核病灶、NTM感染、结核病灶存在的部位等多种因素的影响;T-SPOT.TB检测用于指导结核病诊断时,需综合考虑各项因素。     

Diagnostic evaluation of urinary lipoarabinomannan at an Ethiopian tuberculosis centre

Direct capture enzyme-linked immunosorbent assay (ELISA) for lipoarabinomannan (LAM) was performed on urine samples from 200 tuberculosis (TB) patients and 800 non-TB patients routinely diagnosed among consecutive suspects in an Ethiopian TB centre. 50 healthy Ethiopians, 50 healthy individuals and 100 non-TB patients from Norway served as controls. Of the TB patients, 139 (69.5%) were positive for acid-fast bacilli (AFB). In the remaining cases the diagnosis was based on suggestive clinical findings. All Ethiopian non-TB patients were AFB negative and showed no clinical evidence of TB. In the Ethiopian groups, 148 (74%) of the TB patients, 105 (13.1%) of the non-TB patients and 5 (10%) of the healthy controls were positive by the LAM-ELISA. 113 (81.3%) of AFB positives and 35 (57.4%) of AFB-negative TB patients had positive LAM-ELISA. In the Norwegian groups all were LAM negative. The sensitivity and specificity of the LAM-ELISA for TB patients versus Ethiopian non-TB patients were 74% and 86.9%, respectively; the positive and negative predictive values were 58.5% and 93.0%. This study suggests that detection of LAM in the urine of TB patients may improve case finding and that diagnostic tests based on this principle may serve as valuable supplemental tools in TB control.     

Reducing the number of sputum samples examined and thresholds for positivity: an opportunity to optimise smear microscopy.

SETTING: Urban health clinic, Nairobi. OBJECTIVE: To evaluate the impact on tuberculosis (TB) case detection and laboratory workload of reducing the number of sputum smears examined and thresholds for diagnosing positive smears and positive cases. DESIGN: In this prospective study, three Ziehl-Neelsen stained sputum smears from consecutive pulmonary TB suspects were examined blind. The standard approach (A), > or = 2 positive smears out of 3, using a cut-off of 10 acid-fast bacilli (AFB)/100 high-power fields (HPF), was compared with approaches B, > or = 2 positive smears (> or = 4 AFB/100 HPF) out of 3, one of which is > or = 10 AFB/100 HPF; C, > or = 2 positive smears (> or = 4 AFB/100 HPF) out of 3; D, > or = 1 positive smear (> or = 10 AFB/100 HPF) out of 2; and E, > or = 1 positive smear (> or = 4 AFB/100 HPF) out of 2. The microscopy gold standard was detection of at least one positive smear (> or = 4 AFB/100 HPF) out of 3. RESULTS: Among 644 TB suspects, the alternative approaches detected from 114 (17.7%) (approach B) to 123 cases (19.1%) (approach E) compared to 105 cases (16.3%) for approach A (P < 0.005). Sensitivity ranged between 82.0% (105/128) for A and 96.1% (123/128) for E. The single positive smear approaches reduced the number of smears by 36% compared to approach A. CONCLUSION: Reducing the number of specimens and the positivity threshold to define a positive case increased the sensitivity of microscopy and reduced laboratory workload.     

Phenotypic analysis in peripheral blood lymphocytes of patients with pulmonary tuberculosis]

In order to clarify the immunological aspects of tuberculosis, phenotype of peripheral blood lymphocytes was analyzed in 96 patients with pulmonary tuberculosis and 43 normal controls. In tuberculosis, CD8 positive T cells were significantly decreased in comparison with normal controls and regarding expression of CD3, CD4, CD19, CD16 and gamma delta T cell receptor (TCR), there was no significant difference between tuberculosis and normal controls, though in tuberculosis CD4 positive T cells tended to be high as compared to normal controls. With respect to previous therapeutic condition, the group with previous chemotherapy had significantly high CD4 positive cells, low CD8 positive cells and low gamma delta TCR positive cells as compared to the group without previous chemotherapy. Between groups divided by Type or Expansion of pulmonary tuberculosis without previous chemotherapy, there was no significant difference regarding any surface markers. Regarding pulmonary tuberculosis with previous chemotherapy, the group of Type II had significantly high CD4 positive cells and low CD8 positive cells as compared to Type III, and in the group of Expansion 3, CD19 positive cells were significantly high as compared to Expansion 2. Characteristic of phenotype in peripheral blood lymphocytes of patients with pulmonary tuberculosis is predominant CD4 positive cells in a corresponding decrease in the proportion of CD8 positive cells and is no elevation of gamma delta TCR positive T cells.     

Increased Interleukin-4 production by CD8 and gammadelta T cells in health-care workers is associated with the subsequent development of active tuberculosis

We evaluated immune responses to Mycobacterium tuberculosis in 10 health-care workers (HCWs) and 10 non-HCWs and correlated their immune status with the development of active tuberculosis (TB). Twenty individuals were randomly recruited, tested, and monitored longitudinally for TB presentation. Peripheral blood mononuclear cells (PBMCs) from donors were stimulated with M. tuberculosis and tested for cell proliferation and the production of interferon (IFN)- gamma, interleukin (IL)-5, and IL-4, by use of enzyme-linked immunosorbent or flow-cytometric assays. HCWs had higher levels of cell proliferation (24,258 cpm) and IFN- gamma (6373 pg/mL) to M. tuberculosis than did non-HCWs (cell proliferation, 11,462 cpm; IFN- gamma, 3228 pg/mL). Six of 10 HCWs showed increased median percentages of CD8+IL-4+ (4.7%) and gammadelta +IL-4+ (2.3%) T cells and progressed to active TB. HCWs who remained healthy showed increased median percentages of CD8+IFN- gamma+ (25.0%) and gammadelta +IFN- gamma+ (8.0%) and lower percentages of CD8+IL-4+ (0.05%) and gammadelta +IL-4+ (0.03%) T cells.     

Role of endoscopic ultrasound guided fine needle aspiration/biopsy in the evaluation of intra-abdominal lymphadenopathy due to tuberculosis

BACKGROUNDIntra-abdominal lymphadenopathy due to tuberculosis (TB) poses a diagnostic challenge due to difficulty in tissue acquisition. Although endoscopic ultrasound guided fine needle aspiration/biopsy (EUS-FNA/B) has shown promise in the evaluation of mediastinal lymph nodes, its role in the evaluation of intra-abdominal lymphadenopathy is not clear. AIMTo assess the role of EUS-FNA/B in the evaluation of intra-abdominal lymphadenopathy due to TB.METHODSThis was a retrospective study where patients with intra-abdominal lymphadenopathy who underwent evaluation with EUS-FNA/B were included. TB was diagnosed if the patient had any one of the following: (1) Positive acid fast bacilli (AFB) stain/TB GeneXpert/TB-polymerase chain reaction/AFB culture of tissue sample; and (2) Positive Mantoux test and response to anti-tubercular therapy. EUS-FNA reports, clinical reports and imaging characteristics of patients were recorded for a detailed analysis of patients with TB. RESULTSA total of 149 patients underwent an EUS-FNA/B from lymph nodes (mean age 51 ± 17 years, M:F = 1.2). Benign inflammatory reactive changes were seen in 45 patients (30.2%), while 54 patients (36.2%) showed granulomatous inflammation with/without caseation. Among these, 51 patients (94.4%) were confirmed to have TB as per pre-defined criteria. Patients with TB were more likely to have hypoechoic and matted nodes [40 patients (67.7%)]. EUS-FNA/B was found to have a sensitivity and specificity of 86% and 93% respectively, with a diagnostic accuracy of 88% in the evaluation of intra-abdominal lymphadenopathy due to TB.CONCLUSIONEUS-FNA/B has a high diagnostic yield with a good sensitivity and specificity in the evaluation of intra-abdominal lymphadenopathy due to TB. However, the validity of these findings in populations with low prevalence of TB needs further evaluation.     

Adenosine deaminase estimation and multiplex polymerase chain reaction in diagnosis of extra-pulmonary tuberculosis

SETTING: Extra-pulmonary tuberculosis (EPTB), including mycobacteriosis, contributes 15-20% of all tuberculosis (TB) cases. The diagnosis of EPTB remains elusive because of the inadequate sensitivity of routine and conventional bacteriological methods for the detection of Mycobacterium tuberculosis and related organisms in clinical specimens such as cerebrospinal fluid (CSF), pleural fluid and peritoneal fluid. OBJECTIVE: To develop a better diagnostic marker for EPTB. DESIGN: In our study, 179 cases of EPTB were analysed for acid-fast bacilli (AFB) smear, adenosine deaminase activity (ADA) and multiplex polymerase chain reaction (PCR). Although estimation of ADA is helpful, its sensitivity and specificity varies widely. On the other hand, a multiplex PCR using amplicons such as IS6110, dnaJ gene and hsp65 genes has high sensitivity (60-88%) and specificity (81-100%). RESULTS: On comparing AFB and ADA results with PCR, the PCR is clearly more effective than AFB (P < 0.001) and ADA estimation (P < 0.02) in CSF. The same result was observed with peritoneal fluid (P < 0.001 vs. P < 0.05) and pleural fluid (P < 0.001 and P < 0.05). CONCLUSION: The study shows that multiplex PCR remains the best tool and is a much better marker for diagnosing EPTB.     

应用磁纳米捕获-PCR技术检测痰液结核分枝杆菌的研究

目的采用磁纳米捕获技术富集痰液中的结核分枝杆菌,以提高PCR检测的灵敏度,快速诊断结核病。方法以普通PCR为对照,采用磁纳米捕获技术富集187份结核和非结核呼吸系统疾病患者痰标本中的结核分枝杆菌,进行PCR检测。结果 152份肺结核患者痰标本41份(27.0%)涂片抗酸染色阳性,72份(47.4%)普通PCR检测阳性,126份(82.9%)磁纳米捕获-PCR检测阳性。35份非结核呼吸系统疾病患者,痰标本中抗酸染色均为阴性,1份普通PCR和磁纳米捕获-PCR检测均阳性,该患者临床诊断肺部感染合并陈旧性肺结核。结论采用磁纳米捕获技术富集痰液中的结核分枝杆菌,可显著提高PCR检测的灵敏度。     

Utility of Fine Needle Aspiration Cytology (FNAC) in the diagnosis of tuberculous lymphadenitis compared to GeneXpert in a tertiary health care center in Northern Odisha,India

BackgroundDiagnosis of extrapulmonary tuberculosis including tuberculous lymphadenitis (TBLN) is challenging because of its atypical clinical presentation, paucibacillary nature of mycobacteria at the infected sites, variation in sensitivity of a test to specimens collected by different methods and from different infected tissues.MethodsIn the present study, suspected individuals for lymph node tuberculosis irrespective of age were enrolled prospectively and specimens were collected aseptically by fine needle aspiration (FNA). After the implementation of exclusion criteria, FNA specimens from a total of 278 cases of suspected TBLN were evaluated for cytomorphology (FNAC), presence of acid-fast bacillus (AFB) in smear microscopy and specific detection of mycobacterial DNA in cartridge-based nucleic acid amplification test (CBNAAT).ResultsThe results showed high prevalence of Type II (59.71%), followed by Type I (34.53%) and Type III (5.75%) pattern in FNAC. Non-type II patterns were significantly high in regions outside of the head and neck region (P = 0.031; OR = 2.125) and had an increasing trend of their occurrences with progression of age. The most affected age group was between 16 and 30 years with female preponderance documented in individuals below 45 years, whereas male preponderance was observed in higher age group patients, majority of whom had infected lymph nodes outside of HAN region (P = 0.063, OR = 1.998). The results also showed high sensitivity of CBNAAT (83.04%) method followed by FNAC (72.17%) with AFB smear exhibiting the disappointing results (sensitivity of 10.86%) compared to the CRS. High percentage of positivity was observed in Type III (AFB:25% vs CBNAAT: 100%) followed by Type II (AFB:10.2 vs CBNAAT: 76.5), while low detection was observed from samples with Type I (AFB:4.2 vs CBNAAT: 50). Interestingly, CBNAAT detection of TB was shown to be unaffected by gender, age and site of infection.ConclusionThe study suggests a possible contributary role of age and gender for cytomorphological pattern distribution of TBLN at various body parts. Although FNAC detected TB in 77.1% of cases which were identified positive by CBNAAT and/or AFB, it is being solely based on cytomorphology cannot be used alone as a reliable diagnostic method for TBLN detection. Further, the negative results in CBNAAT for FNAC positive cases may not necessarily be non-TB cases and must be evaluated by other diagnostic modalities. We recommend for both cytomorphological investigation and CBNNAT for the fine needle aspirates from suspected TBLN and subsequent treatment to reduce the disease burden.