| Full text | |
| Author(s): |
De la Torre, David
;
Astolfi-Ferreira, Claudete S.
;
Chacon, Ruy D.
;
Piantino Ferreira, Antonio J.
Total Authors: 4
|
| Document type: | Journal article |
| Source: | VETERINARY SCIENCES; v. 6, n. 1, p. 9-pg., 2019-03-01. |
| Abstract | |
Avian rotavirus A (ARtV-A) is a virus that affects young birds, causing acute diarrhea and economic losses in the poultry industry worldwide. The techniques used for the diagnosis of ARtV-A include electron microscopy, isolation in cell culture, and serology, as well as molecular techniques, such as the reverse transcription-polymerase chain reaction (RT-PCR). The objective of this work was to standardize a real-time RT-polymerase chain reaction (RT-qPCR) using SYBR Green chemistry for the rapid detection and quantification of ARtV-A from bird tissues and materials fixed on FTA cards on the basis of the nucleotide sequence of segment 6 (S6), which codes for the structural VP6 protein of ARtV-A. The results show the efficient amplification of the proposed target, with a limit of detection (LoD) of one copy gene (CG) per microliter of cDNA and a limit of quantification (LoQ) of 10 CGs per microliter. The efficiency of the primers was determined to be 95.66% using a standard curve, with an R-2 value of 0.999 and a slope of -3.43. The specificity was determined using samples coinfected with ARtV-A, the chicken parvovirus, the chicken astrovirus, and the avian nephritis virus as positive controls and commercially available vaccines of the infectious bronchitis virus, infectious bursa disease virus, avian reovirus and healthy organs as negative controls. This technique, which lacks nonspecific PCR products and dimers, demonstrated greater sensitivity and specificity than conventional RT-PCR, and it reduced the analysis time by more than 50%. (AU) | |
| FAPESP's process: | 15/09348-5 - Molecular detection, quantitation, tissue distribution and determination of pathological changes of enteric viruses related to enteric disease in chickens. |
| Grantee: | Luis Fabian Nuñez Naranjo |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 13/08560-5 - Pathogenicity study of chicken enteric viruses (Astrovirus, avian nephritis virus and Parvovirus) in experimentally infected chickens |
| Grantee: | Antonio José Piantino Ferreira |
| Support Opportunities: | Regular Research Grants |