The severe acute respiratory syndrome coronavirus (SARS-CoV-2) pandemic has resulted in significant shortages of RT-PCR testing supplies including RNA extraction kits. The goal of our study was to determine if a simplified heat-RNA release method would provide comparable detection of SARS-CoV-2 without the need for nucleic acid extraction. RT-PCR results using the ChromaCode HDPCR™ SARS-CoV-2 were compared using the heat-RNA release method and an automated RNA extraction system (EMAG). The heat-RNA release method correctly identified 94 % (81/86 nasopharyngeal samples) that were positive for SARS-CoV-2. Five samples that were missed by heat-RNA release method had a mean Ct value: 35 using the automated extraction instrument, indicating a very low viral load. Our findings show that a simple heat-RNA release method is a reasonable alternative for the majority of COVID-19 positive patients and can help overcome the cost and availability issues of RNA extraction reagents.
The analytical performance and cost-effectiveness of the Wampole Toxin A/B EIA, the C. Diff. Quik Chek Complete (CdQCC) (a combined glutamate dehydrogenase antigen/toxin enzyme immunoassay), two RT-PCR assays (Progastro Cd and BD GeneOhm) and a modified two-step algorithm using the CdQCC reflexed to RT-PCR for indeterminate results were compared. The sensitivity of the Wampole Toxin A/B EIA, CdQCC (GDH antigen), BD GeneOhm and Progastro Cd RT-PCR were 85.4%, 95.8%, 100% and 93.8%, respectively. The algorithm provided rapid results for 86% of specimens and the remaining indeterminate results were resolved by RT-PCR, offering the best balance of sensitivity and cost savings per test (algorithm ∼US$13.50/test versus upfront RT-PCR ∼US$26.00/test).
BackgroundUrine is the most frequently cultured specimen type for the majority of clinical microbiology laboratories. Typically, around 30% of cultures are positive for uropathogens with 70% yielding insignificant or mixed growth. BD is developing a software Urine Culture Application (UCA) for the BD Kiestra Total Laboratory Automation (TLA) system to screen images of urine culture plates, sort them based on growth vs. insignificant growth and also allow for presumptive pathogen identification.MethodsDe-identified urine specimens were inoculated onto BD BBL™ CHROMagar™ Orientation Media (CHROM; BD, Sparks, MD), CHROM/Trypticase™ Soy Agar II with 5% Sheep Blood (TSA) biplate, BD BBL MacConkey II agar, and TSA using the BD Kiestra TLA system. Plates were imaged at 24 hours using the BD Kiestra™ ReadA Compact imaging acquisition software and an algorithm was applied to the images using the UCA (Version 2.0). Semi-quantitative measurements of <100, 100–1,000, 1,000–10,000, 10,000–100,000, and >100,000 cfu/mL growth were determined by UCA for all media types and presumptive ID was determined using CHROM. Manual reading of the images by two technologists was the gold standard for comparison. For discrepant results, a third manual reader was used as an arbitrator.ResultsTesting between 877 and 934 urine specimens on each of five media types using UCA resulted in an exact semi-quantitative agreement with manual reading for 85.5–95.0% of specimens (Table 1). If semi-quantitative values ± one category of agreement are included, the number rises to 98.2–99.4% agreement. Using CHROM for presumptive identification of pure or predominant organisms, UCA was in agreement with manual identification in 251 of 272 cultures (92.3%). Of the 21 discrepant organisms, 19 were classified as “other” by manual reading but were identified as specific organisms by UCA. Definitive organism identification was not performed.ConclusionUCA was able to accurately categorize bacterial growth into five semi-quantitative categories using five media types. Pure and predominant uropathogens were accurately identified from CHROM using UCA. The use of UCA software application may enable laboratories to save time screening urine cultures by allowing more efficient use of technologist time. Disclosures All authors: No reported disclosures.
Background Candida auris is an emerging multidrug-resistant pathogen that can persist in the environment and lead to healthcare-associated outbreaks. Residents of long-term acute care hospitals (LTACHs) are at particular risk for carriage of both MDROs and C. auris. However, there are few data on co-colonization rates of C. auris with other MDROs in LTACHs.MethodsWe conducted a point prevalence survey for MDROs, C. auris and C. difficile in a Chicago LTACH in March 2019. A combined axilla/groin E-swab (Copan) was collected and plated for C. auris isolation using CHROMagar Candida (Hardy). A rectal E-swab (Copan) was collected for C difficile PCR and MDRO detection including Carbapenem-resistant Enterobacteriaceae (CRE), Extended-spectrum B-lactamases (ESBLs) and Vancomycin-resistant Enterococci (VRE). Each swab was plated directly on VACC agar (Vancomycin, Amphotericin B, Ceftazidime, Clindamycin) and CHROMagar ESBL (Hardy). Bruker MALDI-TOF was used for bacterial and yeast identification and disc diffusion method for antimicrobial susceptibility testing. ESBL phenotypic confirmation was done using double-disc synergy method per CLSI guidelines. Carbapenemase production was confirmed using Xpert Carba-R assay (Cepheid). C. difficile PCR was performed using Xpert C. difficile/Epi assay (Cepheid).ResultsOf 38 patients 36 were eligible for the study (2 patients declined). Overall, 26/36 (72%) patients had an MDRO. Eight (22%) patients were positive for C. auris. Eight (22%) patients had ESBLs (2 P. mirabilis and 6 E. coli), six (17%) had CREs that were all blaKPC positive (4 K. pneumonia, 1 E. coli, and K. pneumoniae). Eight (22%) patients were positive for other gram-negative (GN)-MDROs including 1 A. baumanii, 3 P. aeruginosa, 2 E. cloacae, 1 E. asburiae and 1 P. aeruginosa, and A. baumanii. 20 patients (56%) had VRE colonization. Five (14%) were C. difficile PCR positive. 7/ 8 (87.5%) patients with C. auris were also colonized with another MDRO (2 VRE, 1 ESBL, 1 VRE, ESBL and KPC, 1 VRE and GN-MDRO, 1 VRE, ESBL and GN-MDRO, 1 VRE, KPC, and GN-MDRO).ConclusionWe found a high rate of MDRO co-colonization among patients with C. auris carriage. Continuous active surveillance may be appropriate in LTACHs to limit the spread of C. auris and other MDROs.Disclosures All authors: No reported disclosures.
A pragmatic, multiphase prospective quality improvement initiative was performed to determine whether a positive displacement connector (PD) causes reduction of central line-associated bloodstream infection (CLABSI), occlusion, and catheter hub colonization when compared with a neutral displacement connector with alcohol disinfecting cap (AC). Patients with an active central vascular access device (CVAD) were enrolled March 2018 to February 2019 (P2) and compared to the prior year (P1). Two hospitals were randomized to use PD without AC (Hospital A) and with AC (Hospital B). Two hospitals utilized a neutral displacement connector with AC (Hospitals C and D). CVADs were monitored for CLABSI, occlusion, and bacterial contamination during P2. Of the 2454 lines in the study, 1049 were cultured. CLABSI decreased in all groups between P1 and P2: Hospital A, 13 (1.1%) to 2 (0.2%); Hospital B, 2 (0.3%) to 0; and Hospitals C and D, 5 (0.5%) to 1 (0.1%). CLABSI reduction was equivalent between P1 and P2 with and without AC, at around 86%. The rate of occlusion per lumen was 14.4%, 12.1%, and 8.5% for Hospitals A, B and C, D, respectively. Hospitals using PD had a higher rate of occlusion than those that did not (P = .003). Lumen contamination with pathogens was 1.5% for Hospitals A and B and 2.1% for Hospitals C and D (P = .38). The rate of CLABSI was reduced with both connectors, and PD reduced infections with and without the use of AC. Both connector types had low-level catheter hub colonization with significant bacteria. The lowest rates of occlusion were found in the group using neutral displacement connectors.
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