RESUMO
The objectives of this study were to assess the ideal volume of Copan FecalSwab™ (FS) preserved stool sample to use with the BD MAX™ Enteric Bacterial Panel and the Extended Enteric Bacterial Panel (BDM GIP) and to compare the performance of FS to the recommended Meridian Para-Pak Cary-Blair medium (PP) for the BDM GIP. Three different input volumes (10, 25, and 50⯵L) of stool inoculated with American Type Culture Collection strains representing the targets detected by BDM GIP were tested. Additionally, 144 unpreserved stool samples submitted for gastrointestinal (GI) testing were transferred to PP and FS media and tested by the BDM GIP using 10⯵L of PP and 50⯵L of FS media. A 100% agreement was observed between PP and FS results. The performance of 50⯵L of FS stool preserved sample was equivalent to 10⯵L of traditional Cary-Blair PP preserved specimens for GI pathogens detection using the BDM GIP.
Assuntos
Fezes/microbiologia , Microbioma Gastrointestinal , Técnicas Microbiológicas/métodos , Manejo de Espécimes/métodos , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Meios de Cultura , Gastroenterite/diagnóstico , Gastroenterite/microbiologia , Microbioma Gastrointestinal/genética , Humanos , Técnicas de Diagnóstico Molecular , Sensibilidade e EspecificidadeRESUMO
The aim of this study was to develop a multiplex PCR test to detect Achromobacter xylosoxidans (AX), Burkholderia cepacia (BC), Pseudomonas aeruginosa (PSA) and Stenotrophomonas maltophilia (SM) directly from CF patient's respiratory samples using the open mode of the BD MAX™ System. A total of 402 CF respiratory samples were evaluated by culture and PCR. Specific sets of primers and probes for each target were designed in-house. Out of 402 samples tested, 196 were identified as negative and 206 as positive by culture for AX, PSA, BC and SM. Among culture positive samples, PCR detected 21/27 AX, 4/5 BC, 138/140 PSA and 29/34 SM. In addition, PCR assay identified 35 samples as positive that were initially negative by culture for those 4 targets. The CF BDM test proved to be an excellent tool to detect AX, BC, PSA and SM by real-time PCR on an automated platform.
Assuntos
Técnicas Bacteriológicas/métodos , Testes Diagnósticos de Rotina/métodos , Infecções Estafilocócicas/diagnóstico , Staphylococcus aureus/classificação , Staphylococcus aureus/isolamento & purificação , Técnicas Bacteriológicas/normas , Testes Diagnósticos de Rotina/normas , Humanos , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Reprodutibilidade dos Testes , Infecções Estafilocócicas/microbiologiaRESUMO
A new multiplex PCR test was designed to detect Mycobacterium chelonae, Mycobacterium abscessus group, and Mycobacterium fortuitum complex on the BD MAX System. A total of 197 clinical samples previously submitted for mycobacterial culture were tested using the new protocol. Samples were first treated with proteinase K, and then each sample was inoculated into the BD MAX Sample Buffer Tube. Extraction and multiplex PCR were performed by the BD MAX System, using the BD MAX ExK TNA-3 extraction kit and BD TNA Master Mix, along with specific in-house designed primers and probes for each target. The limit of detection of each target, as well as specificity, was evaluated. Of 197 clinical samples included in this study, 133 were positive and 60 were negative for mycobacteria by culture, and another 4 negative samples were spiked with M. chelonae ATCC 35752. The new multiplex PCR on the BD MAX had 97% concordant results with culture for M. abscessus group detection, 99% for M. chelonae, and 100% for M. fortuitum complex. The new multiplex PCR test performed on the BD MAX System proved to be a sensitive and specific test to detect M. chelonae, M. abscessus group, and M. fortuitum complex by real-time PCR on an automated sample-in results-out platform.
Assuntos
Reação em Cadeia da Polimerase Multiplex , Infecções por Mycobacterium/diagnóstico , Infecções por Mycobacterium/microbiologia , Mycobacterium chelonae/genética , Mycobacterium fortuitum/genética , Mycobacterium/genética , Reação em Cadeia da Polimerase em Tempo Real , Humanos , Reação em Cadeia da Polimerase Multiplex/instrumentação , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/instrumentação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Group B Streptococcus detection directly from Copan ESwab collected samples, using the BD Max GBS assay, was evaluated on receipt in the laboratory and after 24 h at room temperature. Results were compared to those using Lim broth enrichment PCR and culture. No significant difference was observed between 24 h ESwab and Lim broth PCRs.
Assuntos
Técnicas Bacteriológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Manejo de Espécimes/métodos , Streptococcus agalactiae/isolamento & purificação , Feminino , Humanos , Recém-Nascido , Reação em Cadeia da Polimerase/métodos , Gravidez , Sensibilidade e EspecificidadeRESUMO
A multiplex real-time PCR was validated on the BD Max open system to detect different Mycobacterium tuberculosis complex, Mycobacterium avium complex, and Mycobacterium spp. directly from clinical samples. The PCR results were compared to those with traditional cultures. The multiplex PCR assay was found to be a specific and sensitive method for the rapid detection of mycobacteria directly from clinical specimens.