Review Article Creative Commons, CC-BY
The Use of Primers in PCR Detection of Pathogens of Human and Veterinary Interest and Other Brilliant Ideas.
*Corresponding author: Carlos Navarro Venegas, Associate professor according to FAVET, Faculty of Veterinary and Livestock Sciences (FAVET), University of Chille.
Received: March 06, 2023; Published: April 10, 2023
The incorporation of the fantastic idea of the biochemist and doctor in chemistry Kary Mullis has allowed, along with the design of primers for his protocol, to amplify part of the genome of everything that contains DNA as starting material and in the case of some viruses that contain RNA as genomic material, a previous reaction that synthesizes DNA allows it. A database completes the scene and that’s it.!!! the existence of any suspected pathogen can be validated, regardless of the origin of the sample, animal or human.
The design of primers for a Polymerase Chain Reaction (PCR), the brilliant idea of Kary Mullis, can clearly contribute to the detection of any pathogen that contains DNA or RNA as a genome. The design of primers can be done using various internet platforms, such as Oligoperfect Primer Designer . The PCR protocol is universal since its invention and the other characteristics of the protocol are provided by the computer program.
In short, in human or animal virology, taught in university undergraduate and postgraduate teaching, the specific PCR protocol can now be implemented for the specific detection of the pathogen of interest.
Material and Methods
In our Faculty (FAVET) there is an animal virology laboratory that has a cell culture room. If we take into account that viral isolation continues to be the gold standard or viral detection test, the incorporation of the technique devised by Kary Mullis  together with the design of matches obtained from a computer program such as OligoPerfect complete the idea, prior access to a database: Genbank .
The chosen program provides a primer ranking that is generated by the best combination of thermodynamic parameters, such as GC nucleotide percentage, melting temperature, and obviously nucleotide complementarity, by entering the requested DNA fragment size.
Once the pair of primers of the presented ranking has been chosen, its synthesis is entrusted to a commercial company. Once synthesized and received, all that remains is to program a thermal cycler with the characteristics of the pair of primers and that’s it...!!! everything works perfectly using a conventional PCR system that allows the observation of the DNA fragment through a 2% agarose gel electrophoresis, followed by a photographic record of the evidence using a UV transilluminator.
Discussion and Conclusion
The design of primers that includes the characteristics to be incorporated into a conventional PCR protocol has already been used several times in our laboratory and since 2014 they have been incorporated into the molecular detection of pathogens of veterinary interest using models used that have been made known through the Internet without import the “pedigree” of the journal [4-16].
The foregoing has been reflected in the fact that our country can now count on at least 35 new veterinarians, graduates of our famous House of Studies.
We thank the students of FAVET for making their dreams come true with us and to Dr. Aron Mosnaim, from the Wolf Foundation, Illinois, USA (since 2020).
- (2020) Oligoperfect Primer Designer. Invitrogen®.
- Mullis K, Faloona F (1987) Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods in Enzymology 155: 335-350.
- Genbank® (2023) NIH genetic sequence database.
- Jara P, Céspedes P, Navarro C (2018) Canine Distemper Virus detection based in Hemaglutinine gen as target in reverse transcriptase-Polymerase Chain reaction. IVS p. 1-8.
- Salas V, Pizarro J, Navarro C (2018) Phylogenetic analysis of canine distemper virus detected in Chile. IJCR 10: 72402-72407.
- Gallegos M, Céspedes P, Pizarro J, Navarro C (2018) Is the M gene of Canine Distemper vius a eligible target for detection? EASJALS p. 1-11.
- Méndez-Valenzuela VK, Jara MA, Navarro C (2021) Canine Distemper Virus: Multiple detection of the H and N genes by the Polymerase Chain Reaction associated with Reverse Transcriptase. CJVDS p.1-4.
- Vera C, Jara MA, Navarro C (2022) A preliminary studio for the F gen of Canine Distemper Virus as target for phylogenetic analysis. GSCARR p.1-14.
- Correa V, Céspedes PF, Navarro C (2019) Promising use of Polymerase Chain Reaction Associated to Reverse Transcription for the Detection of the America-1 Lineage of Canine Distemper Virus. IJRSZ p. 1-8.
- Mateo, F., Céspedes, PF., Navarro, C. 2019. The Phosphoprotein Gene from Canine Distemper Virus as Target in Viral Detection. IJZAB p. 1-10.
- Pincheira D, Céspedes P, Pizarro J, Navarro C (2018) Molecular detection of Canine Distemper Virus through the use of the Large Polymerase Gene. EASJALS p. 1-5.
- Navarro C, Muñoz C, Céspedes P (2019) The Nucleocapside Protein Gene As Excellent Target For Detection Of Canine Distemper Virus by Reverse Transcriptase-Polymerase Chain Reaction. Am J Biomed Sci & Res p. 1-7.
- Bolívar P, Céspedes P, Navarro C (2019) Use of the Reverse Transcription-Polymerase Chain reaction for differential detection of two lineages of the canine distemper virus in Chile. IVS p. 1-9.
- Abarca MJ, Hidalgo E, Raggi LA, Navarro C (2018) Genotypic evidence of infection by Canine Distemper Virus in maned Wolf from a zoological collection. IJSR p. 1-10.
- Hidalgo-Hermoso E, Cabello J, Vega C, Kroeger Gómez H, Moreira Arce D, et al. (2020) An eight-year survey for canine distemper virus indicates lack of exposure in the endangered Darwin’s fox (Lycalopex fulvipes). J Wild Dis 56(2): 482-485.
- Vergar Wilson V, Hidalgo Hermoso H, Sánchez C, Abarca M, et al. (2021) Canine Distemper Outbreak by Natural Infection in a Group of Vaccinated Maned Wolves in Captivity. Pathogens