Use of Primer Design to Detect the Glycoprotein C Gene of Canine Herpes Virus by Polymerase Chain Reaction

Abstract

The fundamental difference between a virus and microorganisms lies in the way they generate progeny and some viruses destroy the cell membrane, a phenomenon called cytolysis. The herpes virus has this cytolytic capacity and in veterinary medicine, several important herpes viruses have been described, among which is the Canine Herpes Virus type 1 (CaHV-1) and this study contributed both to the genomic characterization of a national isolate of the CaHV-1 virus -called RP5-as well as to small animal veterinary medicine with a specific, sensitive, and fast diagnostic method result. For this, detection of the CaHV-1 glycoprotein C gene was performed by Polymerase Chain Reaction using a pair of primers designed in silico. Using a temperature gradient thermocycler, the alignment temperature (55ºC) was established by the clear and unequivocal observation of a DNA fragment of approximately 200 bp. These fragments were sequenced and a percentage of nucleotide identity (NIP > 94%) was established with respect to the official GenBank data and indicates that the amplified fragment corresponds to the gC gene fragment of CaHV-1. Thus, this methodology constitutes another technique of choice for the detection of CaHV-1 in Small Animal Medicine.

The Nobel prize and its consequences

“…Of all the winners, Kary Mullis stands out, whose brilliant invention has been part of several final works of Veterinary Medicine students and there are those who mention that the above allows to separate the study of diseases as before and after the PCR [5,6,7,8,9]…”

Some details in the study of viruses that affect to small animals

“…Then, the detection of the glycoproteins gB and gC of CaHV-1 was carried out and in the second case (gC) the design of in silico primers was used, that is, the detection was carried out by means of a PCR reaction with "own" primers [14].…”

Identification of genomes: Clustal Omega and BLAST: One introduction

“…Knowing the foregoing and the existence of the Genbank® database (https://www.ncbi.nlm.nih.gov/genbank/ ), today there are no excuses for the detection of any pathogen or for identifying which genotype (for example) would belong [1,2,3,4].…”