Haptoglobin response in serum and pleural fluid of tuberculin reactor cattle assessed by culture and gross pathology.

Purpose: To report case of extrapulmonary tuberculosis with choroidal tuberculoma as the presenting sign. Method: Case report. Result: A 47-year-old woman with uremia had two choroidal tumors in the left eye. A fundus examination showed the tumors were about 6×4.5×2 mm and 7×5×2 mm in size separately. They were close to each other and located at the nasal and inferior-nasal quadrant of the retina adjacent to the disc. Fluorescence angiography revealed patches of early hypofluorescence and late hyperfluorescence scattered over the area of the two tumors. Indocyanine green angiography, vitreous tuberculosis polymerase chain reaction, and serial ocular sonography were also performed. Chest radiography revealed a right pleural effusion. Computed tomography revealed wall thickening of the ascending colon with multiple regional Lymph nodes and ascites, and two osteolytic lesions at the L1 and L2 vertebrae. Magnetic resonance imaging of thoraco-lumbar spine favored tuberculosis spondylitis at the T9, T12 and L 1-3 vertebral bodies with subligmentous spreading of an abscess. A trace of acid-fast bacilli was found in the L2 spinal aspiration with a positive culture for Mycobacterium tuberculosis complex. After one year of anti-tuberculosis treatment, the choroidal lesions gradually subsided. Conclusion: Choroidal tuberculoma mimicking metastatic malignancy can be a rare presenting sign of systemic tuberculous infection and can be treated with systemic antituberculous medication.

Background: Tuberculosis (TB) remains one of the deadliest communicable diseases. Tuberculosis is caused by Mycobacterium tuberculosis and classified as Pulmonary Tuberculosis (PTB) and Extra Pulmonary Tuberculosis (EPTB). EPTB account for 15-20 % cases of tuberculosis. EPTB diagnosis is challenging due to inadequate sample volume, pauci-bacillary nature and unusual clinical presentation. There are number of tests available for the diagnosis of EPTB but conventional microscopy has low sensitivity and although culture is gold standard method, it takes longer time for yielding positive result. On the other side, GeneXpert due to its rapidity and sensitivity help in early diagnosis and management of tuberculosis. Culture of Mycobacterium Tuberculosis Complex (MTBC) is the accepted reference standard for confirmation of tuberculosis infection and is necessary for Drug Susceptibility Testing (DST). Although solid media has been used for over 100 years, liquid culture media is increasingly being introduced in low and middle income countries for rapid detection of MTBC. To compare the efficacy of solid culture method using Lowenstein Jensen (LJ) media with GeneXpert and liquid culture method using Mycobacterium Growth Indicator Tube (MGIT) 960 for detection of Mycobacterium tuberculosis. Materials and methods: In this cross sectional study 118 suspected EPTB samples including CSF, lymphatic aspirate, pleural fluid, ascitic fluid, synovial fluid, pus were collected. Smear microscopy, culture both in solid media and liquid media using MGIT 960 method and GeneXpert machine were done. Moreover antitubercular drug sensitivity test was done by proportion method in L-J media. Results: Out of 118 samples, GeneXpert detetcs 26 (22.03%) where as the liquid MGIT 960 method detects 27 (22.88%) and solid culture detects 26 (22.03%) positive samples for Mycobacterium tuberculosis. 01 (1.8%) case was detected as multidrug resistant TB. Conclusion: GeneXpert have significant role in diagnosing EPTB patient within shorter period of time. But GeneXpert can detect only rifampicin resistance where as DST in L-J media detects other first line drugs sensitivity too. Liquid culture method using MGIT 960 yields more positive result than solid culture, also liquid culture shows rapid identification of Mycobacterium tuberculosis. Chatt Maa Shi Hosp Med Coll J; Vol.23 (2); July 2024; Page 65-68

Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

Diagnostic and treatment delays increase the severity and transmission of pulmonary tuberculosis (PTB). This study aimed to evaluate TB diagnostic and treatment delays in acid-fast bacilli (AFB) smear-negative patients. This was a retrospective observational study. Patients with positive AFB culture of Mycobacterium tuberculosis complex (MTC) were selected from among hospitalised patients with a diagnosis of pneumonia. Admission ward, anti-tuberculosis treatment and the duration of AFB culture were compared between smear-positive and smear-negative patients. Of the 70 patients with positive isolation of MTC in AFB culture, 27 (38.5%) were smear-negative; of these, 18 (66.7%) were not isolated while in hospital, and 17 (63%) were neither diagnosed nor treated for TB. In contrast, 41 of the 43 smear-positive patients (95.3%) were directly admitted or quickly transferred to the isolation room and started on anti-tuberculosis treatment (P < 0.001). Samples from smear-negative patients required more time to grow MTC in AFB culture than those of smear-positive patients (23 days vs. 14 days, P < 0.001). Diabetes was significantly associated with AFB smear positivity, with an odds ratio of 12.2. Negative AFB smears caused significant diagnostic and treatment delay. Patients staying in the general ward were exposed to TB patients who were not diagnosed in time.

New assay to diagnose and differentiate between Mycobacterium tuberculosis complex and nontuberculous mycobacteria

Surveillance of Tuberculosis in an Acute Care Hospital in the City of Buenos Aires, Argentina

Evaluation of techniques for post-mortem diagnosis of Mycobacterium tuberculosis complex infection in goats.

We conducted a multicenter study to determine whether Mycobacterium tuberculosis complex (MTBC) cultures in automated broth-based systems could reliably be considered negative sooner than 6 weeks. Laboratory sites used Bactec MGIT or BacT/Alert and tracked results of time to detection of all mycobacteria (TTD-all, n = 1547) and of MTBC (TTD-MTBC, n = 466) over 6-month periods from primarily (93%) respiratory specimens. Cumulative percentages by day detected and median TTD of initial and follow-up specimens were analyzed. The median TTD-MTBC for MGIT (n = 6 sites) was 14 days. For laboratories using standard processing procedures, 100% of MTBC were detected from initial and follow-up specimens in 28 and 35 days, respectively, and no yield of MTBC on solid or MGIT liquid media was observed after 5 weeks. The median TTD-MTBC for BacT/Alert (n = 3 sites) was 18 days, with 95% and 100% detected within 37 and 42 days, respectively. Analysis of TTD of positive MTBC cultures in broth can predict the probability of culture negativity at defined time points. Receipt of interim negative reports earlier than 6 weeks could assist clinicians in considering alternative diagnoses and could alter the timing and prioritization of public health interventions. Laboratories should analyze their own TTD data to inform protocol decisions. Laboratories using MGIT could issue reports of no growth of MTBC on initial specimens as early as 4 weeks and for patients undergoing treatment as early as 5 weeks postinoculation.

BackgroundTuberculosis (TB) is a significant global health problem. Nucleic acid amplification tests (NAATs) are valuable in reducing delays to initiation of therapy and infection control protocols. A retrospective study was performed to assess the utilization and performance of a laboratory developed Mycobacterium tuberculosis complex (MTBC) PCR assay (TBPCR) for diagnosis of pulmonary (PTB) and extrapulmonary (EPTB) tuberculosis.MethodsStudy site was a 4 hospital system in suburban Chicago. All culture confirmed TB specimens with complete laboratory data from January 2002 to December 2016 were included. Patient records were accessed using an electronic data warehouse, following approval from Institutional Review Board. Standard microbiology procedures were followed for smear and culture of MTBC. A lab-developed real time PCR targeting a 123 bp region of the IS6110 insertion sequence of MTBC was performed on smear positive specimens or if ordered by physician. Clinical and laboratory data was compared with TBPCR results for all culture confirmed cases.ResultsThere were 151 culture positive patients and 2186 TBPCR performed. Median age of patients at diagnosis was 49 years (IQR 33–66), 74 (49%) were female and 14 were on immunosupressive therapy. The mean number of samples tested per patient was 2. Of culture positive specimens, 59% were from a respiratory source and 3 were MDR; ordering of TBPCR was higher in specimens from PTB source (58.4%) as compared with EPTB source (37%). Combined sensitivity of the TBPCR on all specimen types was 86.6% (95% CI 76.3–93.1); 90.3% for PTB specimens alone (95% CI 78.2–96.4). Specificity was 100% (95% CI 99.5–100), PPV 100% (95% CI 90.5–100%) and NPV 99.5% (95% CI 98.8–99.8%), and were similar for all specimen types. Sensitivity of TBPCR was 97% in smear positive and 79% in smear negative PTB specimens. The median time to culture positivity was 7 days longer in specimens that were TBPCR negative compared with those that were positive (P = 0.14, NS), however, TBPCR shortened time to diagnosis by 13 days.ConclusionWe found TBPCR to be underutilized in both PTB and EPTB although it was found to be a rapid and reliable method for early diagnosis. Education regarding utility of NAATs could be useful in low burden areas where paucibacillary disease is more common, especially in EPTB.Disclosures All authors: No reported disclosures.

Background/Aims: Gastric aspirate specimen is accepted as an alternative specimen of choice to sputum in the diagnosis of childhood pulmonary tuberculosis (TB); and specimen microscopy is usually the only available bacteriologic confirmatory test for streaming cases into the National TB Control Programme treatment protocol. Doubts expressed about the continued relevance of this test among the HIV-infected are based on observations that the bacteriologic yield of acid-fast smears of gastric aspirate specimens from these patients is markedly reduced. This study is aimed at determining the validity of acid-fast smear of gastric aspirates among the HIV-infected and compare with those of the HIV-uninfected. Design, Place and Duration of Study: Diagnostic study. Suspected tuberculosis patients registered in the paediatrics department of University of Benin Teaching Hospital Original Research Article International Journal of TROPICAL DISEASE & Health, 4(1): 111-122, 2014 112 were prospectively recruited from January 2010 to March, 2011. Methodology: A total of 263 children were recruited. Voluntary counseling and testing for HIV was done for each child. Two to three gastric aspirate specimens were also collected from each child using standard gastric aspirate (GA) collection protocols. On each specimen, Zeihl-Neelsen (ZN) staining and culture on Ogawa medium were done. After two or more weeks of incubation, typical acid-fast bacilli isolates on Ogawa medium, which failed to grow on para-nitrobenzoic acid-Ogawa (PNB-Ogawa) media were taken as positive culture for Mycobacterium tuberculosis complex (MTBC). MTBC Culture was taken as the “Reference test” for calculations of sensitivity and specificity. Results: Of the 263 children surveyed 116 (44.0%) were HIV-infected while 147 (56.0%) of them were HIV-uninfected. Among HIV-infected patients, the sensitivity was 38.3% [95%CI:24.4 – 52.2] while the specificity was 95.7% [95% CI:90.9 – 100.0]. Among the HIV-uninfected patients, the sensitivity was 22.0%[95%CI:10.5 – 33.5] while the specificity was 99.0%[95% CI:96.9 -100.0]. The sensitivity of the acid-fast smear in the HIV-infected group of patients was significantly higher than that of the HIV-uninfected group (p = 0.0401).The difference in specificity between the two groups was not statistically significant (p=0.496). There was also no significant difference in their respective positive-predictive values (85.7 vs 91.7)(p = 0.60). Conclusion: The sensitivity of GA acid-fast smear was significantly higher among the HIV infected group.

Role of Xpert MTB/RIF in Bronchoalveolar lavage fluid of sputum-scarce, suspected Pulmonary TB patients

Objective To quantify haptoglobin response to respiratory tract disease in feedlot cattle, and to investigate its ability to predict disease outcome and response to antibiotic treatment. Design Randomized clinical trial. Animals 60 feedlot calves with clinical respiratory tract disease. Procedure Calves were randomly assigned to receive a standard antibiotic treatment regimen (TRT), or to observation pens without antibiotic treatment. Serum haptoglobin concentration was measured at initial and final examinations. Calves were examined for presence of gross pulmonary lesions at slaughter. Results Mean ± SD serum haptoglobin concentration at initial examination was 67 ± 108 mg/dl, with range of 0 to 508 mg/dl. Haptoglobin concentration at initial examination was similar for the TRT group and the group that did not receive antibiotic treatment, but at final examination, TRT-group calves had lower (P &lt; 0.01) mean values. Calves receiving antibiotic treatment had haptoglobin concentration at or near zero at final examination. Calves not receiving antibiotic treatment had only slightly lower mean haptoglobin concentration at final examination, compared with initial examination. Within treatment groups, haptoglobin concentration was similar for cases with different outcomes. Calves with gross pulmonary lesions a: slaughter had numerically higher, although statistically similar, haptoglobin concentrations at initial examination, compared with calves without lesions. Conclusions Feedlot cattle with clinical respiratory tract disease have a large and variable haptoglobin response. Antibiotic treatment resulted in lower serum haptoglobin values, although low values were not required for full clinical recovery. Clinical Relevance Serum haptoglobin concentration may be an indicator of response to antibiotic therapy, although it appears to be unrelated to case severity or need for treatment. (Am J Vet Res 1996; 57:646–649)

Tuberculosis (TB) remains one of the biggest global health challenges. Whilst the greatest burden of active disease is seen in Asia and Africa [1], TB remains a significant issue in the UK. Miliary TB is one of the severest manifestations of TB disease [2–4]. Up-to-date clinicopathological data on miliary TB from the developed world are lacking. We undertook a comprehensive 6-year review (2007–2012) of cases presenting to a single UK centre with an ethnically diverse population with high levels of population exchange with the Indian Subcontinent and Africa. Miliary TB was defined as the presence of miliary nodules on thoracic imaging in patients who presented with symptoms compatible with the diagnosis and either culture of Mycobacterium tuberculosis complex or culture-negative patients with clinical and/or histological features compatible with TB who were commenced on a course of antituberculous therapy (ATT). Miliary TB: high level of CNS involvement best investigated by head MRI which suggests re-evaluation of NICE guidance

Background: Tuberculosis is a highly infectious disease and has the highest burden with it. Diagnosis of tuberculosis in many countries is still dependent on microscopy. For developing countries with a large number of cases and financial constraints, evaluation of rapid and inexpensive diagnostic methods has great importance. Culture of Mycobacterium tuberculosis complex (MtbC) is the accepted reference standard for confirmation of TB infection and is necessary for drug susceptibility testing (DST). There are several methods for culturing MtbC using solid and liquid media. Although solid media has been used for over 100 years, liquid culture media is increasingly being introduced in low and middle income countries (LMIC).&#x0D; Objective: The purpose of the present study was to compare the efficacy of solid culture and liquid culture in the diagnosis of pulmonary tuberculosis.&#x0D; Methodology: This cross sectional study was done in the Department of Microbiology at Sir Salimullah Medical College, Dhaka and National Institute of Chest Disease &amp; Hospital (NIDCH), Dhaka during the period of January 2016 to December 2016 for a period of 1(one) year. Sputum samples from suspected MDR-TB patients were collected by purposive sampling technique from OPD of Sir Salimullah Medical College (SSMC) and NIDCH. Microscopy, liquid culture in liquid MGIT 960 media were done for MTB diagnosis.&#x0D; Result: This study shows the comparison of results of microscopic examination of solid culture and liquid culture (MGIT 960). The liquid MGIT 960 method detected more positive samples than solid culture 68% vs 67%. The mean turnaround time of detection (TTD) of MTB was 34.3±5.2 days for Lowenstein-Jensen media and 17.5±3.8 days for MGIT 960 (p value &lt;0.05). So, liquid culture gave earlier result than solid culture.&#x0D; Conclusion: Liquid culture more positive result than solid culture under microscope in smear of sputum and also liquid culture gave earlier result than solid culture.&#x0D; J Shaheed Suhrawardy Med Coll, June 2019, Vol.11(1); 28-31

Serodiagnosis of tuberculosis and leprosy by enzyme immunoassay.