RESUMEN
BACKGROUND: Apart from major health concerns associated to the SARS-coronavirus-2 (SARS-CoV-2) pandemic, also the diagnostic workflow encountered serious problems. Limited availability of kit components, buffers and even plastics has resulted in suboptimal testing procedures worldwide. Alternative workflows have been implemented to overcome these difficulties. Recently a liquid based sample prep has been launched as solution to overcome limitations in relation to nucleic acid extraction. OBJECTIVE: Multicenter evaluation of the QIAprep& Viral RNA UM kit (QIA P&A) for rapid sample preparation and real-time PCR detection of SARS-CoV-2 in comparison to standardized laboratory testing methods. STUDY DESIGN: Selected samples of the routine diagnostic workflow at Clinical Microbiology Laboratories of four Dutch hospitals have been subjected to the rapid QIA P&A protocol and the results have been compared to routine diagnostic data. RESULTS: Combining results of manual and automated procedures, a total of 377 clinical samples of which 202 had been tested positive with a wide range of CT values, showed almost complete concordance in the QIA P&A assay for samples up to CT values of 33 with one exception of CT 31. Prospectively 60 samples were tested and also showed 100 % concordance with 5 positives. The method has been automated by two centres. CONCLUSIONS: Despite an input of only 8 µL of clinical sample, the QIA P&A kit showed good performance for sample preparation and amplification of SARS-CoV-2 and can contribute as a rapid molecular testing strategy in managing the CoV-2 pandemic.
Asunto(s)
Prueba de COVID-19/métodos , COVID-19/diagnóstico , COVID-19/virología , Tamizaje Masivo/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , SARS-CoV-2/genética , Técnicas de Laboratorio Clínico/métodos , Humanos , Técnicas de Diagnóstico Molecular/métodos , Pandemias/prevención & control , Estudios Prospectivos , Manejo de Especímenes/métodos , Flujo de TrabajoAsunto(s)
Bartonella henselae/inmunología , Enfermedad por Rasguño de Gato/diagnóstico , Anticuerpos Antibacterianos/sangre , Enfermedad por Rasguño de Gato/microbiología , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática/métodos , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Sensibilidad y Especificidad , Pruebas SerológicasAsunto(s)
Bartonella henselae/genética , Sangre/microbiología , Enfermedad por Rasguño de Gato/sangre , Enfermedad por Rasguño de Gato/microbiología , Bartonella henselae/aislamiento & purificación , ADN Bacteriano/sangre , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , Humanos , Linfadenitis/sangre , Linfadenitis/microbiología , Reacción en Cadena de la PolimerasaRESUMEN
Bordetella holmesii is a Gram-negative bacterium first identified in 1995. It can cause pertussis-like symptoms in humans. B. holmesii contains insertion sequences IS481 and IS1001, two frequently used targets in the PCR diagnosis of Bordetella pertussis and Bordetella parapertussis infections. To investigate the prevalence of B. holmesii in Finnish and Dutch patients with pertussis-like symptoms and whether B. holmesii has caused any false-positive results in diagnostic PCRs, B. holmesii-specific real-time PCRs were developed. The Finnish methods were conventional IS481 PCR and B. holmesii-specific real-time PCR (LightCycler, Roche) targeting the B. holmesii recA gene. The Dutch methods were IS481 and IS1001 PCRs with conventional or real-time formats and B. holmesii-specific real-time PCR targeting the homologue of IS1001. Of 11,319 nasopharyngeal swabs, 2804 were collected from Finnish patients from 2000 to 2003, and 8515 from Dutch patients from 1992 to 2003. B. holmesii DNA was not found in the samples analysed. The results suggest that B. holmesii is not among the causative agents of pertussis-like symptoms in Finnish and Dutch patients and thus does not in practice confound IS481 and IS1001 PCRs.
