BackgroundMetallo-β-lactamase (MBL)-producing isolates have a strong impact on diagnostic and therapeutic decisions. A high frequency of MBL-producing gram-negative bacilli has been reported worldwide. The current study was based on determining the incidence of MBL-producing imipenem-resistant clinical isolates and investigating the β-lactamase gene variants in strains conferring resistance to a carbapenem drug (imipenem).MethodsA total of 924 gram negative isolates were recovered from a tertiary care hospital in Lahore, Pakistan, during a two-year period (July 2015 to February 2017). The initial selection of bacterial isolates was based on antibiotic susceptibility testing. Strains resistant to imipenem were processed for the molecular screening of β-lactamase genes. Statistical analysis for risk factor determination was based on age, gender, clinical specimen and type of infection.ResultsThe rate of imipenem resistance was calculated to be 56.51%. Among the 142 strains processed, the phenotypic tests revealed that the incidence of MBLs was 63.38% and 86.61% based on the combination disc test and the modified Hodge test, respectively. The frequencies of blaTEM, blaSHV, blaOXA, blaIMP-1, and blaVIM genes were calculated to be 46%, 34%, 24%, 12.5% and 7%, respectively. The co-expression of blaMBL (blaIMP and blaVIM) and blaESBL (blaTEM, blaSHV, blaOXA) was also detected through multiplex and singleplex PCR. blaOXA, blaTEM and blaSHV coexisted in 82% of the isolates. Co-expression of ESBL and MBL genes was found in 7% of the isolates.ConclusionTo our knowledge, this is the first report from Pakistan presenting the concomitant expression of blaOXA, blaTEM and blaSHV with blaIMP-1 and blaVIM in MBL-producing gram-negative bacilli.

Novel corona virus SARS-CoV-2, causing coronavirus disease 2019 (COVID-19), has become a global health challenge particularly for developing countries like Pakistan where overcrowded cities, inadequate sanitation, little health awareness and poor socioeconomic conditions exist. The SARS-CoV-2 has been known to spread primarily through direct contact and respiratory droplets. However, detection of SARS-CoV-2 in stool and sewage, have raised the possibility of fecal-oral mode of transmission. Currently, quantitative reverse-transcriptase PCR (qRT-PCR) is the only method being used for SARS-CoV-2 detection, which requires expensive instrumentation, dedicated laboratory setup, highly skilled staff, and several hours to report results. Considering the high transmissibility and rapid spread, a robust, sensitive, specific and cheaper assay for rapid SARS-CoV-2 detection is highly needed. Herein, we report a novel colorimetric RT-LAMP assay for naked-eye detection of SARS-COV-2 in clinical as well as sewage samples. Our SARS-CoV-2 RdRp-based LAMP assay could successfully detect the virus RNA in 26/28 (93%) of RT-PCR positive COVID-19 clinical samples with 100% specificity (n = 7) within 20 min. We also tested the effect of various additives on the performance of LAMP assay and found that addition of 1 mg/ml bovine serum albumin (BSA) could increase the sensitivity of assay up to 10 1 copies of target sequence. Moreover, we also successfully applied this assay to detect SARS-CoV-2 in sewage waters collected from those areas of Lahore, a city of Punjab province of Pakistan, declared as virus hotspots by local government. Our optimized LAMP assay could provide a sensitive first tier strategy for SARS-CoV-2 screening and can potentially help diagnostic laboratories in better handling of high sample turnout during pandemic situation. By providing rapid naked-eye SARS-CoV-2 detection in sewage samples, this assay may support pandemic readiness and emergency response to any possible virus outbreaks in future.

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11020 Background: Gender inequality in research funding has been studied extensively; however, the literature lacks evidence in Hematology. We investigated trends in National Institutes of Health (NIH) funding for hematologic malignancies (HM), hematopoietic stem cell transplantation (HSCT), and cellular therapeutics (CT). Methods: The data on Hematology funding was retrieved from NIH Research Portfolio Online Reporting Tools (RePORT) Categorical Spending for fiscal years 2018 and 2019. A total of 6351 entries were reported. Only grants (n=1834) that were related to HM, HSCT, and CT were included. After excluding non-relevant, 975 principal investigators (PIs) were included in the analysis. Additional data regarding PIs was ascertained from the Scopus database, LinkedIn, Doximity, and departmental websites, including the number of publications, number of years of active research, H-index, highest degree, gender, and institution. Data were analyzed using SPSS version 21. Bivariate analyses, using chi-square and t-test, and linear regression analyses were performed. Results: In 2018 and 2019, 1834 grants totaling $799.4 million were awarded by the NIH for malignant hematology research (men 1301, 71% vs women 533, 29%). Of 975 PIs, 680 (70%) were men and 295 (30%) were women. Table highlights gender disparities in NIH funding and associated factors. Most of the grant recipients were Ph.D. or M.D./Ph.D. About 70% of total funding was awarded to male PIs. There were no gender differences in the mean number of grants and mean grant amount. Women had significantly lower years of active research and academic productivity. Conclusions: Although the gender gap in academic hematology has decreased in recent years, the latest trend suggests significant gender inequality in NIH funding for malignant hematology, transplantation, and cellular therapy.[Table: see text]