Mistaken identity of an open reading frame proposed for PCR-based identification of Mycoplasma bovis and the effect of polymorphisms and insertions on assay performance.
J Vet Diagn Invest
; 30(4): 637-641, 2018 Jul.
Article
en En
| MEDLINE
| ID: mdl-29595089
ABSTRACT
Mycoplasma bovis is an important cause of disease in cattle and bison. Because the bacterium requires specialized growth conditions, many diagnostic laboratories routinely use PCR to replace or complement conventional isolation and identification methods. A frequently used target of such assays is the uvrC gene, which has been shown to be highly conserved among isolates. We discovered that a previously described PCR putatively targeting the uvrC gene amplifies a fragment from an adjacent gene predicted to encode a lipoprotein. Comparison of the lipoprotein gene sequence from 211 isolates revealed several single nucleotide polymorphisms, 1 of which falls within a primer-binding sequence. Additionally, 3 isolates from this group were found to have a 1,658-bp transposase gene insertion within the amplified region that leads to a false-negative result. The insertion was not detected in a further 164 isolates. We found no evidence that the nucleotide substitution within the primer-binding region affects the assay sensitivity, performance, or limit of detection. Nonetheless, laboratories utilizing this method for identification of M. bovis should be aware that the region amplified may be prone to nucleotide substitutions and/or insertions relative to the sequence used for its design and that occasional false-negative results may be obtained.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Polimorfismo Genético
/
Reacción en Cadena de la Polimerasa
/
Sistemas de Lectura Abierta
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Mycoplasma bovis
/
Infecciones por Mycoplasma
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
J Vet Diagn Invest
Asunto de la revista:
MEDICINA VETERINARIA
Año:
2018
Tipo del documento:
Article