Direct PCR on Tissue Samples To Detect Mycobacterium tuberculosis Complex: an Alternative to the Bacteriological Culture.

Autor:	Lorente-Leal V; VISAVET Health Surveillance Center, Complutense University of Madrid, Madrid, Spain.; Veterinary Faculty, Animal Health Department, Complutense University of Madrid, Madrid, Spain., Liandris E; VISAVET Health Surveillance Center, Complutense University of Madrid, Madrid, Spain., Pacciarini M; Istituto Zooprofilattico Sperimentale of Lombardia and Emilia-Romagna (IZSLER), National Reference Centre for Bovine tuberculosis, Brescia, Italy., Botelho A; IP National Institute for Agrarian and Veterinary Research (INIAV), Oeiras, Portugal., Kenny K; Central Veterinary Research Laboratory (CVRL), Department of Agriculture, Food and the Marine, Celbridge, Ireland., Loyo B; Laboratorio Regional de Sanidad Animal, Servicio de Sanidad Animal, Dirección General de Producción Agropecuaria, Consejería de Agricultura, Ganadería y Desarrollo Rural de la Junta de Castilla y León, Villaquilambre, Spain., Fernández R; Laboratorio Central de Sanidad Animal. Ministerio de Agricultura, Pesca y Alimentación, Granada, Spain., Bezos J; VISAVET Health Surveillance Center, Complutense University of Madrid, Madrid, Spain.; Veterinary Faculty, Animal Health Department, Complutense University of Madrid, Madrid, Spain., Domínguez L; VISAVET Health Surveillance Center, Complutense University of Madrid, Madrid, Spain.; Veterinary Faculty, Animal Health Department, Complutense University of Madrid, Madrid, Spain., de Juan L; VISAVET Health Surveillance Center, Complutense University of Madrid, Madrid, Spain.; Veterinary Faculty, Animal Health Department, Complutense University of Madrid, Madrid, Spain., Romero B; VISAVET Health Surveillance Center, Complutense University of Madrid, Madrid, Spain bromerom@visavet.ucm.es.
Jazyk:	angličtina
Zdroj:	Journal of clinical microbiology [J Clin Microbiol] 2021 Jan 21; Vol. 59 (2). Date of Electronic Publication: 2021 Jan 21 (Print Publication: 2021).
DOI:	10.1128/JCM.01404-20
Abstrakt:	Bovine tuberculosis (bTB) is an ongoing issue in several countries within the European Union. Microbiological culture is the official confirmation technique for the presence of Mycobacterium tuberculosis complex (MTBC) members in bovine tissues, but several methodological issues, such as moderate sensitivity and long incubation times, require the development of more sensitive and rapid techniques. This study evaluates the analytical and diagnostic performance, comparative to culture, of a real-time PCR targeting the MTBC-specific IS 6110 transposon using a panel of bovine tissue samples sourced from the Spanish bTB eradication campaign. Robustness and repeatability were evaluated in an interlaboratory trial between European Union National Reference Laboratories. The limit of detection with 95% confidence was established at 65 fg/reaction of purified genomic equivalents. Diagnostic sensitivity (Se) and specificity (Sp) were, respectively, 96.45% and 93.66%, and the overall agreement (κ) was 0.88. Cross-reactivity was detected against two mycobacterial isolates identified as Mycobacterium marinum and " Mycobacterium avium subsp. hominissuis ," and whole-genome sequencing (WGS) analysis of the latter isolate revealed an IS 6110 -like sequence with 83% identity. An identical IS-like element was found in other Mycobacterium avium complex species in the NCBI nucleotide and WGS databases. Despite this finding, this methodology is considered a valuable alternative to culture, and the strategy of use should be defined depending on the control or eradication programs. (Copyright © 2021 American Society for Microbiology.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu