The establishment of a recombinase polymerase amplification technique for the detection of mouse poxvirus.
BMC Vet Res
; 19(1): 256, 2023 Dec 06.
Article
em En
| MEDLINE
| ID: mdl-38053140
ABSTRACT
BACKGROUND:
Ectromelia virus (ECTV) is the causative agent of mousepox in mice. In the past century, ECTV was a serious threat to laboratory mouse colonies worldwide. Recombinase polymerase amplification (RPA), which is widely used in virus detection, is an isothermal amplification method.RESULTS:
In this study, a probe-based RPA detection method was established for rapid and sensitive detection of ECTV.Primers were designed for the highly conserved region of the crmD gene, the main core protein of recessive poxvirus, and standard plasmids were constructed. The lowest detection limit of the ECTV RT- RPA assay was 100 copies of DNA mol-ecules per reaction. In addition, the method showed high specificity and did not cross-react with other common mouse viruses.Therefore, the practicability of the RPA method in the field was confirmed by the detection of 135 clinical samples. The real-time RPA assay was very similar to the ECTV real-time PCR assay, with 100% agreement.CONCLUSIONS:
In conclusion, this RPA assay offers a novel alternative for the simple, sensitive, and specific identification of ECTV, especially in low-resource settings.Palavras-chave
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Recombinases
/
Vírus da Ectromelia
Limite:
Animals
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article