Nucleic acid amplification test: Bridging the gap in blood safety & re-evaluation of blood screening for cryptic transfusion-transmitted infection among Indian donors.

Background & objectives: : Nucleic acid amplification test (NAT) in blood donor screening not only detects window period (WP) donors but also those with chronic occult infections which are negative by routine serological screening. This study was conducted to determine the time trend of NAT positivity and seroprevalence of transfusion-transmitted infections (TTIs) through a period of six years and evaluate the strength of NAT as a supplementary test in identifying the cryptic carriers in blood donor population. Methods: : A total of 1,01,411 blood donations were screened between January 2011 and December 2016 by the ELISA and individual donor (ID) NAT Procleix Ultrio Plus Assay. Additional molecular and serological assays were done on the NAT yield samples to differentiate the type of cryptic carriers. Results: : NAT yields comprised 0.05 per cent (50/101411) of the total samples tested with a yield rate of 1/2028. Hepatitis B virus (HBV) contributed to 80 per cent of the total NAT yields and the rest 20 per cent due to hepatitis C virus (HCV). Majority of HBV NAT yields (75%) were from chronic occult donors and 25 per cent were WP donors. Both HBV and HCV NAT yields had a wide range of viral count. There was no HIV NAT yield. A significant decline in the prevalence rate of TTIs through the study period of six years was observed. Interpretation & conclusions: : The cryptic infections found in blood donors increase the risk of TTIs. Blood screening by both serology and NAT can reduce this threat.

Convergence of Minds: For Better Patient Outcome in Intensive Care Unit Infections.

BACKGROUND: There is emergence of resistance to the last-line antibiotics such as carbapenems in Intensive Care Units (ICUs), leaving little effective therapeutic options. Since there are no more newer antibiotics in the armamentarium in the near future, it has become imperative that we harness the interdisciplinary knowledge for the best clinical outcome of the patient. AIMS: The aim of the conference was to utilize the synergies between the clinical microbiologists and critical care specialists for better patient care and clinical outcome. MATERIALS AND METHODS: A combined continuing medical education program (CME) under the aegis of the Indian Association of Medical Microbiologists - Delhi Chapter and the Indian Society of Critical Care Medicine, Delhi and national capital region was organized to share their expertise on the various topics covering epidemiology, diagnosis, management, and prevention of hospital-acquired infections in ICUs. RESULTS: It was agreed that synergy between the clinical microbiologists and critical care medicine is required in understanding the scope of laboratory tests, investigative pathway testing, hospital epidemiology, and optimum use of antibiotics. A consensus on the use of rapid diagnostics such as point-of-care tests, matrix-assisted laser desorption ionization-time of flight mass spectrometry, and molecular tests for the early diagnosis of infectious disease was made. It was agreed that stewardship activities along with hospital infection control practices should be further strengthened for better utilization of the antibiotics. Through this CME, we identified the barriers and actionables for appropriate antimicrobial usage in Indian ICUs. CONCLUSIONS: A close coordination between clinical microbiology and critical care medicine opens up avenues to improve antimicrobial prescription practices.

A ten year analysis of multi-drug resistant blood stream infections caused by Escherichia coli & Klebsiella pneumoniae in a tertiary care hospital.

BACKGROUND & OBJECTIVES: Extensive use of antibiotics has added to the escalation of antibiotic resistance. This study was undertaken to evaluate the association, if any between antibiotic use and resistance in a hospital setting, and also detect the predominant mechanism of antibiotic resistance in Escherichia coli and Klebsiella pneumoniae over a period of 10 years. METHODS: In a retrospective study of 10 years, a total of 77,618 blood culture samples from 2000 to 2009 from indoor patients were screened and those yielding E. coli and K. pneumoniae were included in the study. Antibiotic susceptibility records as well as the percentage of ESBL producers were noted. A total of 423 isolates of 2009 were also screened for AmpC and carbapenemase production. Antibiotic consumption data of 10 years were analysed. RESULTS: ESBL producing E. coli increased from 40 per cent in 2002 to 61 per cent in 2009, similarly there was a significant (P<0.05) rise in resistance to cefotaxime (75 to 97%), piperacillin-tazobactum (55- 84%) and carbapenem (2.4-52%) in K. pneumoniae. A significant (P<0.05) association was observed between resistance and consumption of carbapenem and piperacillin and tazobactum consumption in K. pneumonia. INTERPRETATION & CONCLUSIONS: Our study demonstrated a rise in consumption and resistance to broad spectrum antimicrobial agents and also established an association between consumption and resistance to these antibiotics. Over a period of 10 years, the emergence of pan-resistance in K. pneumoniae could be due to the production of carbapenemases whereas ESBL production was the common mechanism of resistance in E. coli. This study warrants a directed effort towards continued surveillance and antibiotic stewardship to minimize selection pressure and spread.

Monkey Pox arrives in India.

India has confirmed 9 monkey pox (MPXV) cases by the time this editorial is being written. The size of the outbreak clusters is growing each day, as is the geographical spread across international borders. More interestingly it is being recognized as a potent sexually transmitted infection (STI) as it is attributed to close contact with lesions, with no intention of stigmatizing it. This zoonotic virus has a low level of endemicity since decades in west and central Africa, its place of origin. Now since it has spread to 75 countries and more joining, with over 25,539 plus cases, WHO declared it a public health emergency of international concern (PHEIC) on July 23, 2022.

Trend analysis of antimicrobial consumption and development of resistance in non-fermenters in a tertiary care hospital in Delhi, India.

OBJECTIVES: Multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii are becoming increasingly important nosocomial pathogens worldwide. To study the evolution of non-fermenters in a tertiary care hospital, we undertook a retrospective 10 year (1999-2008) trend analysis of antimicrobial consumption and resistance in non-fermenters causing bacteraemia. METHODS: Antibiotic consumption and resistance were analysed by linear regression. The Pearson correlation coefficient was used for assessing correlation between them. RESULTS: A total of 69,010 blood cultures were performed, which grew 15,465 isolates (22% positivity rate), of which 1525 isolates (771 isolates of P. aeruginosa and 754 isolates of A. baumannii) were non-fermenters. Overall antibiotic consumption showed an increasing trend, from 158 to 319 defined daily doses (DDDs)/100 bed-days (r(2)= 0.62, P = 0.007). The largest relative increase in antibiotic consumption was seen for carbapenems (r(2)= 0.68, P = 0.022), followed by ß-lactam/inhibitor combinations (r(2 )= 0.45, P = 0.033), whereas third-generation cephalosporins, fluoroquinolones and aminoglycosides showed no significant changes. A significant increase in resistance in A. baumannii to fluoroquinolones (r(2 )= 0.63, P = 0.006), aminoglycosides (r(2 )= 0.63, P = 0.011) and carbapenems (r(2 )= 0.82, P = 0.013) and in P. aeruginosa to aminoglycosides (r(2 )= 0.59, P = 0.01) was observed. Carbapenem consumption was associated with the development of resistance in A. baumannii (r = 0.756, P = 0.049), whereas no such association was observed for other antimicrobials among non-fermenters. CONCLUSIONS: Our study highlights the growing problem of high antimicrobial consumption. The increasing prevalence of non-fermenters and the emergence of multidrug-resistant A. baumannii are associated with the consumption of carbapenems. The data cannot prove cause and effect.

A prospective study of prevalence of respiratory viruses causing acute respiratory infection in pediatric in-patients during pre- COVID times.

PURPOSE: To find out the prevalence of respiratory viruses causing Acute Respiratory Infection in pediatric in-patients during Pre-COVID times. METHODS: Nasal swabs were collected from children in the age group of 1 month-16 years who were admitted at our hospital with Acute Respiratory Infection. Samples were subjected to nucleic acid extraction and Real time polymerase chain reaction to detect 16 RNA viruses and 2 DNA viruses. The results were interpreted in context of most prevalent viruses detected, their seasonal distribution, co-infecting viruses, co-morbidities in patients with effect thereof and use and effect of antibiotics in those positive for viral etiology. RESULTS: Of the 250 children recruited in the study, viral pathogen was detected in 74% cases. RSV was the most common virus detected with 36.2% positivity (92/254) followed by rhino/entero (19.2%, 49/254), PIV 1,2,3,4 (9.4%, 24/254), Influenza A,B,C (8.2%, 21/254), adenovirus & HBoV (6.2%, 16/254), coronavirus HKU1, NL63, OC43, 229E (4.3%, 11/254), H1N1 (4.7%, 12/254) and hMPV (0.7%, 2/254). Co-infection with 2 or more viruses was seen in 34% cases. Among the cases on whom antibiotics were started, they were withdrawn following test results in 42.3% of the cases. CONCLUSION: The prevalence of viral etiology is high amongst children especially ≤2 years. RSV, rhino/enterovirus, PIV 1,2,3,4 and Influenza virus were more prevalent than others. Rapid, early detection of virus with multiplex PCR will help in early cohorting of the patients thus reducing nosocomial spread of these viruses and prevent injudicious use of antibiotics.

A cross-sectional study of SARS-CoV-2 seroprevalence among asymptomatic healthcare workers in a tertiary healthcare centre: Assessing the impact of PPE guidelines.

PURPOSE: The present study estimates the seroprevalence of SARS-COV-2 among asymptomatic HCWs and assess the impact of various categories of PPE. METHODS: A cross-sectional study of asymptomatic HCW using different levels of PPE as per their risk profile was undertaken between 18th and 24th September 2020. Participant demographics and other relevant details including the levels of PPE used were recorded using a customized questionnaire. IgG antibodies against SARS-COV-2 were detected by chemiluminescence method & used as a surrogate marker for incomplete protection. RESULTS: Out of 1033 HCWs tested, overall SARS-COV-2 sero-prevalence was 25.8%. Univariate and multivariate analysis both demonstrated that ancillary workers including security staff (OR 5.589, P â€‹< â€‹0.001) and sanitary workers (OR 3.946, P â€‹< â€‹0.001) were at significantly higher risk of seropositivity irrespective of the PPE used as per guidelines, whereas doctors were at significantly lower risk of seropositivity (OR 0.307, P â€‹= â€‹0.005). Staff working in office areas was associated with reduced risk of seropositivity (OR 0.21, P â€‹= â€‹0.045). CONCLUSIONS: We document high seroprevalence of SARS-COV-2 antibodies in asymptomatic HCWs. Doctors who are at the highest risk had the lowest seropositivity and seroprevalence among office staff having a risk level comparable to the general community was lower than that reported in general population, supporting the efficacy of PPE practices as per guidelines in these groups. In contrast, much higher rates of seropositivity were seen among ancillary workers despite the availability of adequate PPE. Active screening, proper PPE use as per guidelines, and regular infection control trainings including Covid appropriate behaviour are therefore essential to contain COVID-19 spread among HCW & preventing them to transfer infection to the patients.

Performance of three commercial assays for colistin susceptibility in clinical isolates and Mcr-1 carrying reference strain.

Objective: Commercially available antibiotic susceptibility tests (cAST) for colistin are reported to shows variable performance. The current controversy on the colistin susceptibility testing and scarce data from India has left the clinical laboratories in a dilemma on the appropriate and practical approach to tackle the colistin antimicrobial susceptibility testing (AST) issue. This study was aimed to evaluate the performance of commonly used cAST for colistin against broth microdilution (BMD) as the reference method in the clinical isolates. Materials and Methods: Colistin AST was performed on 225 nonduplicate isolates of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii by BMD as the reference method and compared with Vitek-2, Micronaut-S and E-test. The accuracy of the various cASTs was analysed by assessing categorical and essential agreement (EA). Results: We observed an overall categorical agreement of 98.2%, 99.6% and 96.4% and EA of 92%, 92.4% and 72% for Vitek-2, Micronaut-S and E-test, respectively. Unacceptable rates of major error (10.5%) and very major error (21%) were observed for P. aeruginosa with Vitek-2 and E-test, respectively. All the categorical errors (CEs) (7.7%) with Vitek-2 were seen for minimum inhibitory concentrations ranging within two-fold dilution breakpoint of 2 mg/L. Conclusion: Micronaut-S was found to be an acceptable method for colistin AST. In contrast, E-test was unreliable in terms of EA. Vitek-2 was found to be reliable for colistin AST, although it was more prone to CE near the colistin breakpoints.

Experience of Indian association of medical microbiology external quality assurance scheme centre, New Delhi: Challenges and quality assessment of clinical microbiology laboratories.

Introduction: EQAS program at New Delhi under IAMM was started in January 2014 across North and North east regions of India with 217 participants, which grew up to 540 by 2018. Materials and Methods: In 2014, 4 analytes per year were sent for 3 exercises, i.e. smear culture and serology. 2018 onwards PT analytes were increased from 4 to 12 and comparative performance of techniques analysed. Results: Out of the 22 smears sent for gram staining, ZN staining, Kinyoun staining and Albert staining, completely correct results ranged between 29.55% - 79.9%, 94.3% - 99.2%, 35.5% & 93.8%, respectively. Correct results for culture isolate identification & susceptibility testing and serology exercises varied between 70 & 92.4% and 73.1 & 98.59%, respectively. In the year 2018, 470 responses were received for bacterial culture identification & antibiotic susceptibility testing out of which manual and automated systems were used by 54% & 46% and 52.5% & 47.5% participants, respectively. Techniques used in BBV assays for HBsAg, HCV & HIV found all methods like ELISA, ELFA, CLIA and Card Test performing similarly. The major challenges in running the EQA program included requirement of large amount of specimens for PT item preparation, stability in hot and humid conditions and timely delivery of PT challenges in remote parts of the country. Conclusion: A large number of the participating laboratories (77%) had an overall score of >80% for all exercises, demonstrating acceptable baseline performance of EQAS registered laboratories. However, continued EQAS participation could further improve the quality of results.

Plasma Epstein Barr virus (EBV) DNA as a biomarker for EBV associated Hodgkin lymphoma.

OBJECTIVE: To assess plasma Epstein-Barr virus (EBV) DNA as a biomarker of tumour burden at diagnosis and during therapy in children with Hodgkin lymphoma. DESIGN: Case-control study, with prospective follow-up of the Hodgkin lymphoma cohort (2007-2012). SETTING: Pediatric Hematology Oncology unit of a tertiary care hospital in Delhi. PATIENTS: Thirty children with Hodgkin lymphoma and 70 sex and age-matched controls (benign lymphadenopathy 19, non-lym-phoid malignancy 29, Burkitt lymphoma 5, healthy children 17). INTERVENTION: Positive EBV-staining on immunohistochemistry was defined as EBV-associated Hodgkin lymphoma. Plasma EBV real-time quantitative polymerase chain reaction (PCR) was tested at presentation, after first and last chemotherapy cycles, and on follow-up. MAIN OUTCOME MEASURES: Plasma EBV quantitative PCR was compared between cases and controls. Its kinetics was assessed during and after chemotherapy. RESULTS: EBV quantitative PCR was positive in 19 (63%) Hodgkin lymphoma cases (range 500 to 430,000 copies/mL), with 87.5% accuracy (kappa=0.69) as compared with EBV immunohistochemistry. Sensitivity and specificity of the quantitative PCR were 87.5% and 81.8%, respectively. Only boys showed positive EBV immunohistochemistry and,or quantitative PCR positivity. All controls were quantitative PCR negative. All quantitative PCR positive cases with follow up blood sample showed EBV clearance after the first cycle. A quantitative PCR negative case in long-term remission became positive at relapse. EBV status did not influence survival. CONCLUSION: Plasma EBV-DNA, detectable in EBV-associated Hodgkin lymphoma, becomes undetectable early after initiating therapy. It can be used as a biomarker of treatment response in EBV-associated Hodgkin lymphoma.