Evaluation of Two Real-Time, TaqMan Reverse Transcription-PCR Assays for Detection of Rabies Virus in Circulating Variants from Argentina: Influence of Sequence Variation
| Autor: | Yu Li, Daniel Cisterna, María A Lombardo, Leila Martínez, Cristina Lema, María S Sabio, Paula Becker, Diego A. Caraballo, Fernando J. Beltrán |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Rabies 030106 microbiology lcsh:QR1-502 Argentina diagnostic Biology medicine.disease_cause Real-Time Polymerase Chain Reaction phylogeny Genome lcsh:Microbiology Article 03 medical and health sciences Virology Chiroptera TaqMan medicine Animals Humans rabies virus Amino Acid Sequence Horses Sequence variation Geography Medical nucleoprotein gene Gene Reverse Transcriptase Polymerase Chain Reaction Rabies virus Genetic Variation real time RT-PCR medicine.disease Nucleoprotein Reverse transcription polymerase chain reaction Phylogeography 030104 developmental biology Infectious Diseases Nucleoproteins RNA Viral Cattle |
| Zdroj: | Viruses Volume 13 Issue 1 Viruses, Vol 13, Iss 23, p 23 (2021) |
| ISSN: | 1999-4915 |
| DOI: | 10.3390/v13010023 |
| Popis: | In rabies diagnosis, it is essential to count on a rapid test to give a quick response. The combined sensitivity and robustness of the TaqMan RT-PCR assays (qRT-PCR) have made these methods a valuable alternative for rabies virus (RABV) detection. We conducted a study to compare the applicability of two widely used qRT-PCR assays targeting the nucleoprotein gene (LysGT1 assay) and leader sequences (LN34 qRT-PCR assay) of RABV genomes, in all variants circulating in Argentina. A total of 44 samples obtained from bats, dogs, cattle, and horses, that were previously tested for rabies by FAT and conventional RT-PCR, were used in the study. All variants were successfully detected by the pan-lyssavirus LN34 qRT-PCR assay. The LysGT1 assay failed to detect three bat-related variants. We further sequenced the region targeted by LysGT1 and demonstrated that the presence of three or more mismatches with respect to the primers and probe sequences precludes viral detection. We conclude that the LysGT1 assay is prone to yield variant-dependent false-negative test results, and in consequence, the LN34 assay would ensure more effective detection of RABV in Argentina. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
| Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |