Identification of endogenous Coccidioides posadasii contamination of commercial primary rhesus monkey kidney cells.

Here we describe the identification of endogenous Coccidioides posadasii contamination in commercial rhesus monkey kidney (RhMK) cells and the subsequent nationwide alert that reduced the risk of laboratory exposure. This extraordinary event highlights the necessity for laboratories to remain vigilant in the use of appropriate biosafety procedures, particularly when working with unknown pathogens.

Evaluation of multiple test methods for the detection of the novel 2009 influenza A (H1N1) during the New York City outbreak.

BACKGROUND: In response to the novel influenza A H1N1 outbreak in the NY City area, 6090 patient samples were submitted over a 5-week period for a total of 14,114 viral diagnostic tests, including rapid antigen, direct immunofluorescence (DFA), viral culture and PCR. Little was known about the performance of the assays for the detection of novel H1N1 in the background of seasonal H1N1, H3N2 and other circulating respiratory viruses. In addition, subtyping influenza A became critical for the identification of high risk and/or hospitalized patients with novel H1N1 infection and for monitoring the spread of the outbreak. STUDY DESIGN: This study analyzed the performances of the BinaxNOW Influenza A&B test (BinaxNOW), the 3M Rapid Detection Flu A+B test (3MA+B), direct immunofluorescence, R-Mix culture and the Luminex xTAG Respiratory Virus Panel (RVP) for the detection of seasonal influenza, novel H1N1 and other respiratory viruses. RVP was also evaluated for its ability to differentiate seasonal H1N1, H3N2 and novel H1N1. RESULTS: The sensitivities, specificities, PPVs and NPVs for the detection of novel H1N1, determined by comparing all four-test methods, were: rapid antigen: 17.8%, 93.6%, 77.4%, 47.9%; DFA: 46.7%, 94.5%, 91.3%, 58.9%; R-Mix culture: 88.9%, 100%, 100%, 87.9%; RVP: 97.8%, 100%, 100%, 97.3%. The individual sensitivities of BinaxNOW and 3MA+B as compared to R-Mix culture for the detection of novel H1N1 were 9.6% and 40%, respectively. All unsubtypeable influenza A specimens identified by RVP and tested with the CDC novel H1N1 specific RT-PCR assay were confirmed to be novel H1N1. CONCLUSIONS: Rapid antigen tests, DFA, R-Mix culture and the xTAG RVP test all detected the novel H1N1 strain, but with highly varied sensitivity. The RVP test provided the best diagnostic option as RVP demonstrated superior sensitivity for the detection of all influenza strains, including the novel H1N1, provided accurate influenza A subtyping and identified a significant number of additional respiratory pathogens.

Clinical performance of the 3M Rapid Detection Flu A+B Test compared to R-Mix culture, DFA and BinaxNOW Influenza A&B Test.

BACKGROUND: The rapid diagnosis of influenza allows for prompt patient management and the initiation of appropriate infection control measures to reduce spread in healthcare settings. OBJECTIVE: To evaluate the clinical performance of the 3M Rapid Detection Flu A+B Test (3MA+B) as compared to R-Mix cell culture, direct immunofluorescence assay (DFA) and the BinaxNOW A&B Influenza Test (BinaxNOW). STUDY DESIGN: Five hundred fresh respiratory samples, collected from patients aged 5 days to 99 years with respiratory symptoms, were tested by R-Mix culture, DFA, 3MA+B and BinaxNOW. Analytical sensitivity of 3MA+B was compared to BinaxNOW using replicates of serially diluted clinical samples positive for influenza A or B. RESULTS: Sensitivity, specificity, PPV and NPV for the detection of influenza A and B, respectively, were for R-Mix (96.9%, 100%, 100%, 99.3%; 98.1%, 100%, 100%, 99.8%), DFA (80.4%, 99.2%, 96.1%, 95.3%; 74%, 100%, 100%, 97%), 3MA+B (70.1%, 99.8%, 98.6%, 93%; 86.5%, 98.7%, 88.2%, 98.4%) and BinaxNOW (46.4%, 100%, 100%, 88.6%; 34.6%, 100%, 100%, 93%). R-Mix, DFA and 3MA+B were significantly (P

Clinical evaluation of NucliSENS magnetic extraction and NucliSENS analyte-specific reagents for real-time detection of human metapneumovirus in pediatric respiratory specimens.

In this study, we evaluated the NucliSENS miniMAG (MM) and easyMAG (EM) nucleic acid extraction platforms (bioMérieux, Durham, NC) in combination with the NucliSENS EasyQ basic kit and analyte-specific reagents (ASRs) (bioMérieux) for the detection of human metapneumovirus (hMPV) in respiratory samples. Total nucleic acids from pediatric clinical samples (n = 653) and an hMPV-specific inhibition control (h-IC) were coextracted using the MM and/or the EM. Nucleic acid sequence-based amplification and real-time molecular beacon detection of hMPV were performed using a NucliSENS EasyQ analyzer (bioMérieux). Positive results were confirmed using an in-house-validated reverse transcriptase PCR ASR-based assay. The inclusion of the h-IC monitored the entire process, including the efficiency of nucleic acid extraction, amplification, and detection. The percentages of samples with inhibited amplification of the h-IC after initial NA extraction by EM and MM were 1.88% and 3.17%, respectively. After reprocessing of a new aliquot, the final h-IC inhibition rates were 0% (EM) and 1.06% (MM). The limit of detection of the assay was between 2 (EM extraction) and 10 (MM extraction) RNA copies/reaction, and specificity was 100% when testing viral respiratory isolates and clinical samples. hMPV was detected in 5.6% of pediatric samples tested and was also detected in three coinfections with respiratory syncytial virus (RSV). hMPV was the second most frequently detected respiratory virus in children of 0 to 2 years of age, after RSV. In summary, NucliSENS extraction and ASRs provided a sensitive and specific method for the detection of hMPV in respiratory samples.

Survival of spumavirus, a primate retrovirus, in laboratory media and water.

The persistence of a previously characterized spumavirus strain (strain SV-522) was investigated utilizing various laboratory media and waters, including Eagle's minimal essential medium (EMEM) plus 0% fetal bovine serum (EMEM-0%), EMEM-2%, EMEM-10%, Chlamydia transport medium (CTM), phosphate-buffered saline, distilled, estuarine, and marine water, human serum, and the germicides, ethyl alcohol (70%) and sodium hypochlorite (10%). Experiments were performed at 4 degrees C and/or 23 degrees C. Infectivity endpoints were determined in stock aliquots upon initiation of testing and then after 3, 5, 7, and 10 days. The virus was reisolated from all diluents after 5 days at 23 degrees C and in EMEM-10% after 7 days. The virus was detected in CTM, EMEM-2%, EMEM-10%, and estuarine and marine waters after 7 days at 4 degrees C. Differences in the persistence of the virus may be ascribed to temperature and organic load. Water ionic strengths (e.g., estuarine vs. marine water) had no effect on modifying persistence of viral particles. Infectivity of spumavirus was undetectable after 30 s in 70% ethanol or 10% sodium hypochlorite. After 30 min at 23 degrees C, spumavirus infectivity in normal but not heat-inactivated human serum increased by almost 100-fold. Persistence of infectivity of primate spumavirus after 7 days in media and waters, and the agent's infectious potential in the human host, emphasize a need for cautious recognition during the manipulation of primate cells/organs and in the handling of primates themselves.