Nontuberculous mycobacteria (NTM) represent an increasingly significant cause of pulmonary and extrapulmonary infections, but are sometimes misinterpreted as tuberculosis (TB) owing to overlapping clinical and microbiological characteristics. Conventional diagnostic approaches, such as Ziehl-Neelsen staining and culture in a Mycobacterial Growth Indicator Tube (MGIT) system, are constrained by extended incubation times, are insufficient for accurate species differentiation, and are limited by prolonged incubation periods. Recent molecular and genomic advances have transformed NTM diagnostics by enabling rapid, specific, and high-resolution identification. Line probe assays (e.g., GenoType Mycobacterium CM/AS assay) and multiplex PCR have enhanced the ability to distinguish between NTM species such as Mycobacterium absessus, M. fortuitum, and M. avium complex and M. tuberculosis complex, which is essential for proper treatment and epidemiological mapping. Among newer proteomic platforms, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry has emerged as a transformative, cost-effective technology capable of identifying Mycobacterium species directly from culture isolates through protein fingerprinting. It provides rapid, reproducible, and highly discriminatory identification between closely related species. Next-generation sequencing (NGS) and whole genome sequencing approaches now offer unprecedented insight into species identification, strain typing, and drug-resistance prediction, complementing traditional culture-based susceptibility testing. Newer techniques such as metagenomics NGS (mNGS), targeted NGS (tNGS) multilocus sequence typing, and mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTR) genotyping facilitate subspecies-level resolution and real-time outbreak surveillance. Moreover, molecular beacons, insertion sequence analysis, and repetitive sequence-based polymerase chain reaction (Rep-PCR) enhance detection sensitivity even in paucibacillary samples. The integration of genomic data with automated diagnostic system promises earlier intervention, accurate species delineation, and improved patient outcome.

Non-tuberculous mycobacteria (NTM) are environmental opportunistic pathogens causing chronic pulmonary disease. NTM pulmonary disease (NTM-PD) and bronchiectasis exhibit a bidirectional pathogenic relationship that is particularly under recognized in South Asian regions with high tuberculosis (TB) burden. This review article covers most of the current evidence on the epidemiology, clinical spectrum, pathogenesis, therapeutic advances of NTM-associated bronchiectasis. A well-structured literature search was conducted across PubMed, Web of Science, Scopus, and Google Scholar for articles published within from 2008 to 2025. These studies were specifically focused on NTM -associated bronchiectasis for host pathogen interactions, diagnostic strategies, and treatment outcomes. The prevalence of NTM related bronchiectasis especially with species like Mycobacterium avium complex and Mycobacterium abscessus which were more frequent pathogens all over the world. Its diagnostic dilemma with TB remains widespread due to limited lab capacity and lack of exact species identification, which leads to late or wrong diagnosis. Integration of molecular diagnostic tools, inclusion of NTM within national TB programs, and establishment of regional reference laboratories are essential to improve early detection, targeted treatment, and disease surveillance for bronchiectasis.

Acute respiratory infections (ARIs) are a leading cause of death in children under five globally. The seasonal trends and profiles of respiratory viruses vary by region and season. Due to limited information and the population’s vulnerability, we conducted the hospital-based surveillance of respiratory viruses in Eastern Uttar Pradesh. Throat and nasal swabs were collected from outpatients and inpatients in the Department of Paediatrics, Baba Raghav Das (BRD) Medical College, Gorakhpur, between May 2022 and April 2023. A total of 943 samples from children aged 1 to 60 months were tested using multiplex real-time PCR for respiratory viruses in cases of ARI and SARI. Out of 943 samples tested, the highest positivity was found for parainfluenza virus [105 (11.13%) PIV-1 (79), PIV-2 (18), PIV-4 (18)], followed by adenovirus [82 (8.7%), RSV-B, [68 (7.21%)], influenza-A [46(4.9%): H1N1 = 29, H3N2 = 14), SARS CoV-2 [28 (3%)], hMPV [13(1.4%), RSV-A [4 (0.42%), and influenza-B (Victoria lineage) 1 (0.10%). The maximum positivity of respiratory viruses was seen in children between 1 to 12 months. The wide variation in prevalence of these respiratory viruses was seen in different seasons. This study enhances understanding of the seasonal and clinical trends of respiratory virus circulation and co-infections in Eastern Uttar Pradesh. The findings highlight the importance of targeted interventions to reduce the burden of respiratory infections in this region.

Abstract Guillain–Barré syndrome (GBS) is a rare condition, potentially fatal, immune-mediated disease of the peripheral nerves and nerve roots that leads to weakness, numbness or paralysis. The exact cause of GBS is unknown but usually triggered by different infections (Campylobacter, influenza virus, cytomegalovirus and Epstein–Barr virus, etc.). The incidence of GBS can therefore increase during the outbreaks of infectious diseases such as recently seen in Pune, Maharashtra. Diagnosis and management of GBS can be complicated as its clinical presentation and disease course are variable and unpredictable, because there are no well clinical guidelines available. A universal applicable guideline based on the current literature and expert opinion to assist doctors for the diagnosing and management of GBS. Guideline cover the early identification and diagnosis of GBS, admission to the intensive care unit, treatment indication and selection, monitoring and treatment during progression of disease, prediction of clinical course, outcome and management of complications and sequelae.

BACKGROUND:Tuberculosis (TB), caused by Mycobacterium tuberculosis complex, results in approximately 1.5 million annual deaths globally. Diagnosing extrapulmonary TB (EPTB) remains challenging due to the invasive nature of sample collection and limitations in conventional diagnostic sensitivity. This study evaluates the diagnostic performance of Xpert®Mycobacterium tuberculosis/Rifampicin (MTB/RIF), a nucleic acid amplification test, against direct microscopy for EPTB specimens. In addition, we compare the detection of first-line anti-tubercular drug resistance between Xpert® MTB/RIF and the MTB-DR plus line probe assay. METHODS:From January 2022, to April 2023, 2839 clinically suspected EPTB specimens were collected from patients referred to tertiary care hospitals in Gorakhpur, India. Specimens included lymph node aspirates, pleural fluid, cerebrospinal fluid, and tissue biopsies, processed according to the Indian National Tuberculosis Elimination Program protocols. Diagnostic evaluations employed microscopy (acid-fast bacilli staining), Xpert® MTB/RIF, and MTB-DR plus assays. RESULTS:Of 2839 specimens, Xpert® MTB/RIF detected M. tuberculosis in 339 cases (11.9%), significantly outperforming microscopy (183 cases, 6.4%). The highest positivity rates occurred in tissue biopsies and lymph node aspirates (29%), while genitourinary TB was least frequent. Rifampicin resistance was identified in 14 cases (4.13%), all confirmed as multidrug-resistant TB (MDR-TB) by MTB-DR plus. CONCLUSION:Xpert® MTB/RIF demonstrated superior sensitivity over microscopy, supporting its utility for EPTB diagnosis in low-resource settings. The high MDR-TB prevalence (4.13%) underscores the need for rapid molecular diagnostics to guide treatment. However, global EPTB burden estimates remain inconsistent, necessitating standardized surveillance and diagnostic protocols to improve detection accuracy and inform public health strategies.

The emergence of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant TB (XDR-TB) complicates control efforts. We investigated the prevalence and genetic patterns of pre-XDR and XDRTB, by employing second-line line probe assays (SL-LPA). This prospective cohort study was conducted at the Intermediate Reference Laboratory, from November 2023 to May 2024. We screened 1253 samples via Genxpert followed by florescence microscopy, and mutational/resistance analysis via the GenoType MTBDRplus/FL-LPA (first line-line probe assay) and MTBDRsl/SL-LPA for the mutation patterns. We identified 355 rifampicin drug-resistant TB isolates, 260 of which were smear positive samples obtained for FL-LPA and SL-LPA. Finally, 255 MDR/RR-TB patients were analyzed and 73(28.62

Tuberculosis remains a significant global health challenge, with the re-emergence of Mycobacterium tuberculosis (MTB) posing serious concerns, especially in its extra-pulmonary forms. Rapid and accurate diagnosis is crucial for effective management. This study evaluates the performance of the Xpert MTB/RIF Ultra assay compared with the Xpert MTB/RIF and traditional microscopy (Ziehl-Neelsen stain) in diagnosing extra-pulmonary tuberculosis (EPTB) in a tertiary care center in northern India. A total of 100 samples collected from September 2024 to February 2025 underwent testing using these methods. Our findings indicate that the Xpert MTB/RIF Ultra assay demonstrated superior sensitivity and reliability compared with the Xpert MTB/RIF, whereas microscopy was the least effective. This study is the first of its kind in India, highlighting the enhanced diagnostic capabilities of the Xpert MTB/RIF Ultra assay for EPTB. IMPORTANCE This is the first study to come out of India where the newly launched Xpert Ultra/RIF test has been used to detect Paucibacillary EPTB cases in comparison to the old Xpert MTB/RIF test.

Introduction:Drug-resistant tuberculosis (DR-TB) poses a significant public health threat, with molecular diagnostics playing a pivotal role in understanding the genetic mechanisms of resistance. This study focuses on the patterns of genetic mutations observed in DR-TB cases, with the aim to identify key mutations associated with resistance to rifampicin (RIF) and isoniazid (INH). Methodology:A total of 6,954 non-duplicate clinical samples were obtained from individuals of all age groups, categorized as TB and DR-TB, from seven linked districts between June 2022 and May 2024. The samples were transported under cold chain conditions to an intermediate reference laboratory. TB was confirmed using fluorescence microscopy, and 1,998 sputum-positive samples were analyzed using line probe assay for characterization of genetic mutations. Results:Among the analyzed cases, a total of 136 cases of DR-TB were identified. This included 57 cases (41.92%) of multidrug-resistant TB (MDR-TB), 73 cases (53.68%) of INH monoresistance, and 6 cases (4.4%) of RIF monoresistance. The analysis revealed a high prevalence of rpoB MUT3 (S531L) mutations in 52 cases (82.25%), which is associated with RIF resistance. In high-level INH (katG gene mutation) resistance noted in 83 (63.35%) cases, katG MUT1 (S315T1) was predominant, while low-level INH resistance (inhA gene mutation), inhA MUT1 (C-15T) mutation, was found in 29 (22.13%) cases. Maharajganj and Deoria reported the highest prevalence of rpoB MUT3 (S531L) mutations, while Kushinagar and Sant Kabir Nagar exhibited higher rates of katG MUT1 (S315T1) mutations. Other regions showed notable distribution of rpoB, katG, and inhA gene mutations. Conclusion:The high prevalence of mutations such as rpoB MUT3 (S531L) and katG MUT1 (S315T1) highlights the need for integrating molecular tools into routine workflows to identify genetic mutations. District-specific mutations emphasize the influence of local epidemiological factors on resistance patterns, necessitating region-specific interventions. Continuing research into regional resistance trends are vital to addressing the global DR-TB burden effectively.

Introduction: Acute viral hepatitis (AVH) is mostly caused by hepatitis A virus (HAV) and hepatitis E virus (HEV). Hepatitis B virus, on the other hand, is a major cause of chronic liver disease in children and adolescents. Objectives: To determine the seroprevalence and clinical outcomes of HAV and HEV in children of different age groups in a tertiary care hospital in Eastern Uttar Pradesh, India.Method: During the study period, 382 clinically suspected cases of acute hepatitis were examined. Data included demographic information; and clinical history of illness specifically linked to the hepato-biliary system.Results: A total of 382 samples was screened for AVH of which 58 (15.1%) were HAV positive, 21 (5.4%) were HEV positive and 12 (3.1%) were positive for both anti‑HAV and anti‑HEV antibodies indicating HAV‑HEV co‑infection. There was also significant co-infection between hepatitis A and E. Male children (54%) were more prone to all kinds of viral hepatitis than female children (46%), with the most common age group being 0-5 years in HAV cases and 11-15 years in HEV cases. Icterus was the most prevalent clinical sign in the hepatitis profile, followed by fever, hepatomegaly and nausea.Conclusions: In this tertiary care hospital in Eastern Uttar Pradesh, India, the most frequent viral hepatitis infection in children was hepatitis A, followed by HEV. There was significant co-infection between hepatitis A and E.

Globally, drug-resistant tuberculosis is major public health issue. Multidrug-resistant tuberculosis (MDR-TB), especially resistant to first line drugs (isoniazid [INH] and rifampicin [RMP]), is one of the serious health concerns in India. As per annual India TB report 2022, 2.7% of total TB patients were infected with MDR-TB. Line probe assay (LPA)/GenoTypeMTBDRplus technique based on polymerase chain reaction is a rapid method for the detection of RMP and INH simultaneously. This cross-sectional prospective study includes 405 samples from suspected TB patients from 30.12.2021 to 13.05.2022. Samples were received from linked districts with Intermediate Reference Laboratory (IRL) for fluorescent microscopy (FM) and LPA testing through proper cold-chain under NTEP program and then subjected to FM. Only smear positive sample were subjected to LPA as per manufacturer instructions. Out of 405, a total of 261 samples were smear positive (64.4%). Among 261 samples processed for LPA, 255(97.7%) showed valid result and 6(2.3%) were found invalid. In LPA result, 227 (89.01%) were susceptible to both INH and RMP, 9(3.5%) had MDR-TB, 18(7.05%) shows INH mono resistance and 1(0.3%) shows RMP mono resistance. Patients with MDR-TB were prominent in younger age group (Mean±S.D; 29.6±5.7 vs. 36.6±17.8 years, p value = 0.004). Prevalence of MDR-TB cases is higher in our study than cases reported in annual TB report (2022) of India. LPA is proved highly accurate in rapid detection RMP resistant/MDR-TB cases. Early diagnosis may potentially enable earlier commencement of appropriate TB drugs and the reduction of transmission of MDR-TB and may possibly leading to the complete eradication of TB in India.

Background:The coronavirus disease 2019 (COVID-19) vaccination program among adults in India is one of the highly successful vaccination drives globally, but the acceptance of the COVID-19 vaccine among parents for their children is largely unknown. As parents are primary decision-makers for their children, we aimed to assess parents' perceptions and acceptance for COVID-19 vaccination for children in India. Aim:To assess parental perceptions and acceptance for COVID-19 vaccination for their children (aged <18 years). Materials and Methods:A cross-sectional anonymous web-based survey was designed and conducted from November 1 to December 15, 2022. All the healthcare workers of a tertiary care institute in eastern Uttar Pradesh, having a child aged <18 years, constituted the study population and were invited. They were recruited through snowball sampling and were sent the study questionnaire in Google form through email and WhatsApp. Bivariate analysis was performed to determine the predictors of child vaccination acceptance among the parents. Results:A total of 388 healthcare worker parents (HCWPs) having a child <18 years of age completed the survey. The mean age of the parents was 40.28 ± 11.34 years. The majority (96.9%) of them had already received the recommended two doses of the COVID-19 vaccine. Around 91% of the parents agreed that COVID-19 vaccines are important for children's health, whereas 89% agreed that COVID-19 vaccines are effective. A total of 356 (91.7%) parents were willing to vaccinate their children with a COVID-19 vaccine, of which 91.3% intended to do so as soon as possible. The factors significantly associated with the stated willingness for child vaccination among the participants were younger age (P = 0.008), female gender (P = 0000), currently married status (P = 0.00001), urban residence (P = 0.01), lower monthly income (P = 0.03), absence of any chronic illness (P = 0.0003), history of COVID-19 in the child (P = 0.004), and the child being up-to-date with the routine childhood vaccines (P = 0.01). Conclusion:The HCWPs from eastern Uttar Pradesh, India, have a positive attitude and low vaccine hesitancy toward COVID-19 vaccination for children, with around 92% currently willing to vaccinate their children with COVID vaccine. Further longitudinal studies are required to assess the trend of parental acceptance of COVID vaccine.

Aims: To study the impact of past COVID 19 infections on various hematological parameters and morbidity in patients with active dengue infection by assessing days of hospital stay, platelet transfusion etc. Study Design: Prospective study. Place and Duration of Study: Department of Pathology, Baba Raghav Das Medical college, Gorakhpur, India between June 2022 to November 2022. Methodology: 189 patients with active dengue infection, proven on Dengue ELISA for IgM detection, were included in this prospective study. Hematological parameters along with demographic profiles and clinicopathological details were recorded for the patients at the time of admission. Results: The mean age was comparable between the two groups, dengue patients with history of COVID 19 (Group A) being 26.93±14.27 years and dengue patients with no history of COVID 19 (Group B) being 28.52±17.65 years. Group A patients were found to have higher mean platelet count [68200.00±28153.33] than Group B patients [54181.21±31792.06]. The fatality rate was 2.68% in the Group A patients and 5% in the group B. Conclusion: Our research indicates that individuals with a history of COVID-19 infection have reduced dengue mortality. However, we cannot presume a causal association between dengue and COVID-19 infection prognosis.

Background and Objectives: In India, it is estimated that there are 40 million people suffering from Hepatitis B virus (HBV). Quantification of the viral burden is an important laboratory tool in the management. However, widespread use of different HBV-DNA assays is still affected by the high cost and variable diagnostic precision. The present study was conducted to evaluate the diagnostic precision and co-relation of ALT levels with HBV-DNA by Truenat®-PCR. Materials and Methods: In this prospective cross-sectional study a total of 567 serums were collected from patients by rapid HBsAg, and processed for liver function tests (LFT). The viral HBV-DNA amplification detection was carried out through by Truenat®-PCR test. Results: Out of 567 samples, 452 samples were found to be positive by both rapid and Truenat®-PCR and 106 were negative for HBV-DNA followed by 9 invalid. High ALT level found in 73% of positive patients who had HBV-DNA level (>100000 copies/ml) which is significantly higher in 447 patients as compared to those have below ≤100000 copies/ml. Conclusion: Truenat®-PCR technique is a highly sensitive and can be performed with low resources for effective control of HBV infection. Evaluation of HBV-DNA levels and serum ALT levels showed a significant proportion of patient harbored ongoing viral replication and disease progression.

Background: The ability of Staphylococcus aureus to maintain a clinically very indolent state depends on various lifestyles the bacteria evolved as Small Colony Variants [SCV] phenotype to safeguard themselves causes chronicity of the infection, providing resistance to the host challenge and antibiotics. Aim: A prospective study has been conducted on none-duplicated S. aureus isolates from bloodstream infections were collected from January 2022-December 2022. Methods: We conducted prospective study considering the characteristics morphology, identification etc, antimicrobial resistance patterns of SCVs were also investigated as per standard guidelines. Results: A total of 585 positive blood culture samples were collected with growth of S. aureus. Among which 140 [23.93%] isolates came out to be SCV, remaining 445 [76.06%] were Normal Colony Variants [NCV] of S. aureus. The prevalence of MRSA among 140 SCV came out to be 80 [57.14%] as compared with 445 NCV was 182 [40.89%]. In this study majority with type 2 DM [51.79%], prolonged hospitalization [42.22%], and placement of central line [36.41%]. Most of the MRSA isolates were found among SCV [57.14%], followed by NCV [40.89%]. The mortality rate was 7.69%, in which the majority of the patients belong to NCV, [84.44%] followed by SCV [15.55%]. Conclusion: The SCV phenotype and antibiotic resistance are interlinked; we still need to learn more about the clinical consequences of infection as well as the risk factors for SCV MRSA. These results highlight the importance of SCV infection and support the routine use of clinical laboratory tests that are sensitive for SCV.