[IFN-gamma enzyme-linked immunospot assay versus PPD tuberculin skin test in the diagnosis of tuberculous epididymitis].

OBJECTIVE: To explore the potential application of IFN-gamma enzyme-linked immunospot (ELISPOT) assay in the diagnosis of tuberculous epididymitis (TE) by comparing ELISPOT assay with the traditional purified protein derivative (PPD) tuberculin skin test. METHODS: We examined 13 TE patients using an in-house ELISPOT kit, another 11 TE patients by PPD skin testing, and 57 healthy male volunteers by parallel test with both the methods. RESULTS: Twelve (92.3%) of the 13 TE cases were positive on ELISPOT assay, and 10 (90.9%) of the 11 TE cases positive on PPD skin test, with no statistically significant differences between the two groups (P > 0.05). Among the 57 healthy male volunteers, 8 (14.0%) were positive on ELISPOT, and 28 (49.1%) positive on PPD test, the latter significantly higher than the former (P < 0.001). CONCLUSION: In terms of sensitivity, ELISPOT assay is similar to PPD test in the examination of tuberculous epididymitis. As for specificity, ELISPOT assay seems better than PPD test in differentiating tuberculous epididymitis patients from healthy males.

Improvement cues of lesion absorption using the adjuvant therapy of traditional Chinese medicine Qinbudan tablet for retreatment pulmonary tuberculosis with standard anti-tuberculosis regimen.

BACKGROUND: China is the second highest pulmonary tuberculosis (PTB) burden country worldwide. However, retreatment of PTB has often developed resistance to at least one of the four first-line anti-TB drugs. The cure rate (approximately 50.0-73.3%) and management of retreatment of PTB in China needs to be improved. Qinbudan decoction has been widely used to treat PTB in China since the 1960s. Previously clinical studies have shown that the Qinbudan tablet (QBDT) promoted sputum-culture negative conversion and lesion absorption. However, powerful evidence from a randomized controlled clinical trial is lacking. Therefore, the aim of this study was to compare the efficacy and safety of QBDT as an adjunct therapy for retreatment of PTB. METHODS: We conducted a multicenter, randomized, double-blind, placebo-controlled clinical trial in China. People diagnosed with PTB were enrolled who received previous anti-TB treatment from April 2011 to March 2013. The treatment group received an anti-TB regimen and QBDT, and the control group was administered an anti-TB regimen plus placebo. Anti-TB treatment options included isoniazid, rifampicin, pyrazinamide, ethambutol, streptomycin for 2 months (2HRZES), followed by isoniazid, rifampicin, ethambutol for 6 months (6HRE), daily for 8 months. Primary outcome was sputum-culture conversion using the MGIT 960 liquid medium method. Secondary outcomes included lung lesion absorption and cavity closure. Adverse events and reactions were observed after treatment. A structured questionnaire was used to record demographic information and clinical symptoms of all subjects. Data analysis was performed by SPSS 25.0 software in the full analysis set (FAS) population. RESULTS: One hundred eighty-one cases of retreatment PTB were randomly divided into two groups: the placebo group (88 cases) and the QBDT group (93 cases). A total of 166 patients completed the trial and 15 patients lost to follow-up. The culture conversion rate of the QBDT group and placebo group did not show a noticeable improvement by using the covariate sites to correct the rate differences (79.6% vs 69.3%; rate difference = 0.10, 95% confidence interval (CI): - 0.02-0.23; F = 2.48, P = 0.12) after treatment. A significant 16.6% increase in lesion absorption was observed in the QBDT group when compared with the placebo group (67.7% vs 51.1%; rate difference = 0.17, 95% CI: 0.02-0.31; χ2 = 5.56, P = 0.02). The intervention and placebo group did not differ in terms of cavity closure (25.5% vs 21.1%; rate difference = 0.04, 95% CI: - 0.21-0.12; χ2 = 0.27, P = 0.60). Two patients who received chemotherapy and combined QBDT reported pruritus/nausea and vomiting. CONCLUSIONS: No significant improvement in culture conversion was observed for retreatment PTB with traditional Chinese medicine plus standard anti-TB regimen. However, QBDT as an adjunct therapy significantly promoted lesion absorption, thereby reducing lung injury due to Mycobacterium tuberculosis infection. TRIAL REGISTRATION: This trial is registered at ClinicalTrials.gov, NCT02313610.

[Expression of Mycobacterium tuberculosis early secretory antigenic target-6 protein and its application in detection of Mycobacterium tuberculosis antigen-specific interferon-gamma response].

OBJECTIVE: To obtain the purified early secretory antigenic target-6 (ESAT-6) protein and to evaluate its application in detection of Mycobacterium tuberculosis antigen-specific interferon-gamma (IFN-gamma) response. METHODS: ESAT-6 protein was expressed by genetic engineering. The antigen specificity and reactivity of ESAT-6 were evaluated by Western blot. Using ESAT-6 as the antigen, the antigen-specific IFN-gamma response in patients with tuberculosis, healthy medical workers, and village residents was detected by the Elispot method. The results were also compared with those obtained by a commercial kit (QuantiFERON-TB-GOLD, QFT-G). RESULTS: ESAT-6 protein was successfully expressed and purified, and the antigen specificity of ESAT-6 was confirmed by its recognition by the antigen-specific antibody (anti-ESAT-6). The specificity and sensitivity of the Elispot assay using ESAT-6 as the antigen in detecting the IFN-gamma response was comparable with those of the commercial kit (QFT-G). The positive rates of the Elispot assay for patients with tuberculosis, healthy medical workers and villagers were 36/49 (73.5%), 11/62 (17.7%), and 17/194 (8.8%), respectively, while the rates of the OFT-G method for patients with tuberculosis and healthy medical workers were 38/49 (77.6%) and 14/58 (24.1%), respectively. The sensitivity (73.5%, 77.6%; chi2 = 0.381, P > 0.05) and specificity (82.3%, 75.9%; chi2 = 0.406, P > 0.05) of these two methods did not differ significantly. CONCLUSIONS: Recombinant ESAT-6 protein was expressed and purified. Elispot using recombinant ESAT-6 protein as antigen showed high sensitivity and specificity for detection of Mycobacterium tuberculosis antigen-specific IFN-gamma response. The purified ESAT-6 can be used for diagnosis of Mycobacterium tuberculosis infection.

Indicators for prediction of Mycobacterium tuberculosis positivity detected with bronchoalveolar lavage fluid.

BACKGROUND: The diagnosis of active pulmonary tuberculosis (TB) remains a challenge in clinic, especially for sputum negative pulmonary TB. Bronchoalveolar lavage fluid (BALF) has higher sensitivity than sputum for detection of Mycobacterium tuberculosis (Mtb). However, bronchoscopy is invasive and costly, and not suitable for all patients. In order to make TB patients get more benefit from BALF for diagnosis, we explore which indicator might be used to optimize the choice of bronchoscopy. METHODS: A total of 1539 sputum-smear-negative pulmonary TB suspects who underwent bronchoscopy were recruited for evaluation. The sensitivity, specificity and accuracy of Mtb detection in sputum and BALF were compared. Odds ratios and 95% confidence intervals were used to assess variables that associated with positive acid-fast bacilli (AFB) smear, Mtb culture and nucleic acid amplification test (NAAT) of BALF in sputum-negative and non-sputum-producing pulmonary TB suspects. RESULTS: BALF has significantly higher sensitivity (63.4%) than sputum (43.5%) for Mtb detection by culture and NAAT. 19.7% (122/620) sputum-negative and 40.0% (163/408) non-sputum-producing suspects had positive bacteriological results in BALF. Among sputum-negative and non-sputum-producing pulmonary TB suspects, the positivity of Mtb detection in BALF is associated with a younger age, the presence of pulmonary cavities and a positive result of interferon-gamma release assay (IGRA). Sputum-negative patients under 35 years old with positive IGRA and pulmonary cavity had 84.8% positivity of Mtb in BALF. CONCLUSIONS: Our study indicated that combination of age, the presence of pulmonary cavity, and the result of IGRA is useful to predict the positivity of Mtb detection in BALF among sputum-negative and non-sputum producing pulmonary TB suspects. Those who are under 35 years old, positive for the presence of pulmonary cavity and IGRA, should undergo bronchoscopy to collect BAFL for Mtb tests, as they have the highest possibility to get bacteriologically confirmation of TB.

[Study on specific cellular immunity of enhance intracellular survival antigen of Mycobacterium tuberculosis in patients with pulmonary tuberculosis].

OBJECTIVE: To investigate the level of specific cellular immunity in patients with active pulmonary tuberculosis and its potential relationship with severity of the disease. METHODS: Thirty active pulmonary tuberculosis patients with positive tubercle bacilli in sputum were enrolled for the study. Immune responses including lymphocytes proliferation enhanced by enhance intracellular survival (EIS) antigen and cytokine production including interferon-gamma (IFN-gamma) and interleukin-10 (IL-10), were assayed. Cell proliferation was determined by methyl thiazolyl tetrazolium (MTT), while cytokine production was quantified by enzyme linked immunoadsorbent assay (ELISA). The results were compared to those of 20 healthy individuals and 16 persons recovered from tuberculosis. RESULTS: Cell proliferation response and IFN-gamma production were significantly higher in patients convalescent from tuberculosis compared to patients with active pulmonary tuberculosis, EIS antigen was found to elicit a dominant Th2 cytokine response. CONCLUSION: Impaired Th1 immune response to EIS is observed in patients with active pulmonary tuberculosis. Induction of imbalance of Th1/Th2 immune response may be the main action of EIS, which may be a factor of pathogenesis of tuberculosis.

[Evaluation of the effect of chronic virus infection on laboratory tests results in patients with osteoarticular tuberculosis].

OBJECTIVE: To evaluate the effect of chronic virus infection on laboratory tests results in patients with osteoarticular tuberculosis. METHODS: A total of 121 patients with osteoarticular tuberculosis, who were hospitalized in Shenzhen Third People's Hospital during June 2008 to June 2012, were recruited for analysis. Clinical laboratory tests results were collected for comparison between patients with or without chronic co-infection with virus. RESULTS: Among the 121 patients, thirty patients were co-infected with hepatitis B virus (HBV), two were with Human immunodeficiency virus (HIV), and one was co-infected with HBV, HIV and hepatitis C virus (HCV). Compared to patients with osteoarticular tuberculosis without HBV/HCV/HIV infection, patients with chronic HBV/HCV/HIV virus infection had similar positive rate of laboratory tests including tissue smear acid-fast bacilli (AFB) staining, tissue Mycobacterium tuberculosis (Mtb) culture, tissue Mtb DNA detection, serological test of antibodies against Mtb, and Mtb. antigen-specific interferon-gamma release assay. Similar results were also found for erythrocyte sedimentation rate, C-reative protein level and liver function including Alanine aminotransferase and Aspartate Aminotransferase. CONCLUSION: Chronic infection with HBV/HCV in patients with have no obvious effect on clinical laboratory tests related to tuberculosis.

Multi-site abdominal tuberculosis mimics malignancy on 18F-FDG PET/CT: report of three cases.

18F-fluorodeoxyglucose positron emission/computed tomography (18F-FDG PET/CT) imaging, an established procedure for evaluation of malignancy, shows an increased 18F-FDG uptake in inflammatory conditions. We present three patients with abdominal pain and weight loss. Conventional imaging studies indicated that abdominal neoplasm and 18F-FDG PET/CT for assessment of malignancy showed multiple lesions with intense 18F-FDG uptake in abdomen of the three cases. However, the three patients were finally diagnosed with multi-site abdominal tuberculosis (TB). Of them, two were diagnosed with TB by pathology, one was diagnosed with TB clinically. They recovered after anti-TB therapy. Few reports on accumulation of 18F-FDG in abdominal TB are available in the literature. A high index of suspicion is necessary to achieve an early diagnosis and a better outcome of the disease.

[Role of nitric oxide--NF-kappaB signaling pathway in differentiation of human naive T lymphocytes into Th1/Th2 cells].

AIM: To explore the role of nitric oxide (NO)--NF-kappaB signaling pathway in the differentiation of human naive T lymphocytes into Th1/Th2 cells. METHODS: Human naive T lymphocytes were isolated from umbilical blood. Various concentrations of NO donor sodium nitroprusside (SNP), NO inhibitor NAME and NF-kappaB inhibitor PDTC were added to the culture medium to induce the differentiation of naive T cells towards Th1/Th2 cells. The expressions of intracellular cytokine IFN-gamma and IL-4 were detected by flow cytometry. RESULTS: The treatment of SNP, NAME and PDTC made no difference on the percentage of cells expressing IFN-gamma (Th1) or IL-4 (Th2) in comparison with that of the control group (P < 0.05). CONCLUSION: Signaling pathway of NO--NF-kappaB had no effect on differentiation of human naive T lymphocytes into Th1 and Th2 cells. The role of NO--NF-kappaB signaling pathway in the regulation of types 1 and 2 cytokines may occur mainly at the level of mature Th cells.