Diagnosis of pleural tuberculosis by multi-targeted loop-mediated isothermal amplification assay based on SYBR Green I reaction: comparison with GeneXpert® MTB/RIF assay.

BACKGROUND: Diagnosis of pleural tuberculosis (TB) is tedious owing to its close resemblance with malignant pleural effusion and sparse bacterial load in clinical specimens. There is an immediate need to design a rapid and dependable diagnostic test to prevent unnecessary morbidity/mortality. RESEARCH DESIGN AND METHODS: A multi-targeted loop-mediated isothermal amplification (MT-LAMP) was deliberated using mpt64 and IS6110 to diagnose pleural TB within pleural fluids/biopsies. MT-LAMP products were analyzed by gel-based and visual detection methods, viz. SYBR Green I, SYBR Green I+deoxyuridine triphosphate uracil-N-glycosylase (dUTP-UNG), and dry methyl green reactions. RESULTS: In a pilot study, while assessing pleural TB/non-TB control subjects (n = 40), both SYBR Green I+dUTP-UNG/gel-based MT-LAMP assays exhibited better sensitivity/specificity than SYBR Green I and dry methyl green MT-LAMP. Since it is facile to work with SYBR Green I+dUTP-UNG than gel-based MT-LAMP, we validated the performance of SYBR Green I+dUTP-UNG in a higher number of specimens (n = 97), which revealed somewhat higher sensitivity (85.2 vs. 81.5%) and specificity (97.7 vs. 90.7%) than SYBR Green I MT-LAMP. Furthermore, the sensitivity attained by SYBR Green I+dUTP-UNG MT-LAMP was significantly higher (p < 0.001) than GeneXpert. CONCLUSIONS: Our SYBR Green I+dUTP-UNG MT-LAMP is a simple and reliable method to diagnose pleural TB, which may translate into a point-of-care test.

Predictors of post COVID complications in patients admitted with moderate to severe COVID symptoms: A single center, prospective, observational study.

While the world was still busy battling active COVID-19 infections, a large subset of patients started showing prolonged symptoms or developing complications following an initial recovery from COVID-19. Post covid complications range from mild symptoms such as fatigue, headache, shortness of breath to serious, life threatening conditions like opportunistic infections, deep venous thrombosis, pulmonary embolism, pneumothorax and lung fibrosis. A single center, prospective, observational study was carried out in a tertiary respiratory care institute in North India from June 2021 to August 2021 where 224 cases of previously treated COVID-19/ongoing symptomatic COVID-19 (those patients who were manifesting symptoms beyond 4 weeks), were enrolled and followed up for a period of 3 months to estimate the prevalence of persistent symptoms, complications and any risk factors associated with it. Data analysis was done using SPSS software version 21. Univariate and multivariate analysis done among risk factors and outcome variables. ROC was done on predictor variables and area under curve (AUC) calculated. p value less than 0.05 was considered significant. Among the 24.6% symptomatic patients at follow up, the most common symptom was fatigue (51.8%) followed by dyspnea (43.8%) and anxiety (43.3%). Among the complications of COVID-19, the most common according to our study was fibrosis (15.2%), followed by pulmonary thromboembolism (PTE) (12.1%), echocardiographic abnormalities (11.2%) and pulmonary mucormycosis (5.4%). Female gender, presence of comorbidities, requirement of non-invasive or invasive ventilation during hospital stay emerged as independent risk factors for complications following COVID-19. This study brings forth the huge morbidity burden that COVID-19 brought upon seemingly cured individuals and lists the risk factors associated with persistence of symptoms and complications. This would help to better streamline health resources and standardize follow up guidance of COVID-19 patients.

Insight into diagnosis of pleural tuberculosis with special focus on nucleic acid amplification tests.

INTRODUCTION: Pleural tuberculosis (TB) is the archetype of extrapulmonary TB (EPTB), which mainly affects the pleural space and leads to exudative pleural effusion. Diagnosis of pleural TB is a difficult task predominantly due to atypical clinical presentations and sparse bacillary load in clinical specimens. AREA COVERED: We reviewed the current literature on the globally existing conventional/latest modalities for diagnosing pleural TB. Bacteriological examination (smear/culture), tuberculin skin testing/interferon-γ release assays, biochemical testing, imaging and histopathological/cytological examination are the main modalities. Moreover, nucleic acid amplification tests (NAATs), i.e. loop-mediated isothermal amplification, PCR/multiplex-PCR, nested-PCR, real-time PCR and GeneXpert® MTB/RIF are being utilized. Currently, GeneXpert Ultra, Truenat MTBTM, detection of circulating Mycobacterium tuberculosis (Mtb) cell-free DNA by NAATs, aptamer-linked immobilized sorbent assay and immuno-PCR (I-PCR) have also been exploited. EXPERT OPINION: Routine tests are not adequate for effective pleural TB diagnosis. The latest molecular/immunological tests as discussed above, and the other tools, i.e. real-time I-PCR/nanoparticle-based I-PCR and identification of Mtb biomarkers within urinary/serum extracellular vesicles being utilized for pulmonary TB and other EPTB types may also be explored to diagnose pleural TB. Reliable diagnosis and early therapy would reduce the serious complications associated with pleural TB, i.e. TB empyema, pleural fibrosis, etc.

How familiar are our doctors towards Rabies prophylaxis- A study from coastal south India.

BACKGROUND: Rabies, a 100% fatal disease claims more than 59,000 human lives every year globally. One human life is lost every 15 minutes due to this deadly preventable disease. Timely initiation of post exposure prophylaxis following an animal exposure can result in 100% preventability of this fatal disease. METHODOLOGY: This facility based study was conducted among clinical fraternities of teaching hospitals. A semi structured questionnaire was used for collection of data. Institutional Ethics Committee approval was sought. The study investigators visited the workplace of the participants and distributed the questionnaire. SPSS Ver 16 (Chicago, IL, USA) was used to analyse the data. FINDINGS: Most of the participants knew that veterinary groups and zoo-keepers should be given pre-exposure prophylaxis. Many participants knew about the Intra Muscular schedule of anti-rabies vaccine and its site of administration for pre exposure prophylaxis. It was observed that most participants had knowledge regarding correct intramuscular regimen of anti-rabies vaccine for post-exposure prophylaxis but less than half were able to differentiate between the intramuscular and intradermal regimens. Less than half of participants were aware of the fact that local administration of anti-rabies serum is useful. CONCLUSION: The knowledge regarding WHO categorisation of animal exposure and recommended post exposure prophylaxis according to type of exposure observed to be minimal among clinical fraternity.