How To Detect a Pathogen of Human or Veterinary Interest?

Introduction

Molecular virology, in addition to being a dynamic discipline, has had to incorporate various methodologies that have appeared lately, such as the fantastic idea of Kary Mullis and other scientists. Although this has made it possible to expand the knowledge of viral genomes (RNA or DNA) in terms of size, length or nucleotide sequence thanks to the existence of the Genbank® [1] and some computer programs that allow both the design of primers for a reaction of PCR, the alignment of nucleotide sequences or the determination of the identity of the pathogen of interest. To anyone within this area, the existence of these programs should be unknown, even online and for free. The molecular detection of the pathogenic agent, therefore, is today a task without pretexts, that is, there is no pretext for its implantation anywhere on our planet.

Material and Methods

Today, the existence of the Genbank® has allowed access to many of the nucleotide sequences of various viral agents that affect both humans and other animals. With this valuable information and the famous invention of Kary Mullis [2], a conventional Polymerase Chain Reaction is formed when complemented with 2% agarose gel electrophoresis. Subsequently, by sending the amplified fragment for sequencing and determining the nucleotide identity using other internet platforms such as CLUSTAL Omega [3] and BLAST [4], the detection of the suspected agent can be validated. In the context of the samples to be used for the detection of the agent (human or animal), we have been able to access fragments of organs or fluids from which the nucleic acid involved is extracted. Subsequently, the PCR can be developed by designing the primers and subsequent sequencing in companies dedicated to this mission such as Macrogen Ltd. Finally, thanks to CLUSTAL Omega and BLAST it is possible to align the delivered sequences and subsequently establish the nucleotide identity of the amplified fragment.

Discussion

The indicated methodologies and the existence of free access computer programs have been mentioned in various publications in mainstream journals [5,6].and in others of equal importance and it has been possible to magnify the importance of the molecular study of pathogens of interest and in our case, taking as models those mentioned [7-17].

Conclusion

Thanks to the use of these methodology, at least 35 new veterinarians have graduated in our country, who have incorporated these techniques into their professional knowledge, which are here to stay.

Acknowledgments

We thank the students of FAVET for making their dreams come true with us and Dr. Aron Mosnaim, from the Wolf Foundation, Illinois, USA (since 2020).

Conflict of Interest

No conflict of interest.

References

• Genbank® (2023) NIH genetic sequence database.
• Mullis K, Faloona F (1987) Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods in Enzymology 155: 335-350.
• (2023) CLUSTAL OMEGA. Multiple Sequence Alignment.
• BLAST (2023) Basic Local Alignment Search Tool.
• Hidalgo Hermoso E, Cabello J, Vega C, Kroeger Gómez H, Moreira Arce, 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.
• Vergara Wilson V, Hidalgo Hermoso H, Sánchez C, Abarca M, Navarro C, et al. (2021) Canine Distemper Outbreak by Natural Infection in a Group of Vaccinated Maned Wolves in Captivity. Pathogens 10(1): 51.
• Salas V, Pizarro J, Navarro C (2018) Phylogenetic analysis of canine distemper virus detected in Chile. IJCR 10: 72402-72407.
• Jara P, Céspedes P, Navarro C (2018) Canine Distemper Virus detection based in Hemaglutinine gen as target in reverse transcriptase-Polymerase Chain reaction. IVS 2: 034-041.
• Gallegos M, Céspedes P, Pizarro J, Navarro C (2018) Is the M gene of Canine Distemper virus a eligible target for detection? EASJALS 1(1).
• Vera C, Jara MA, Navarro C (2022) A preliminary studio for the F gen of Canine Distemper Virus as target for phylogenetic analysis. GSCARR. 10(01): 105-118.
• 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 2(1).
• 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 5(1): 18-25.
• Mateo F, Céspedes PF, Navarro C (2019) The Phosphoprotein Gene from Canine Distemper Virus as Target in Viral Detection. IJZAB 2(6).
• 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 5(4): 309-315.
• 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 3: 005-013.
• Pincheira D, Céspedes P, Pizarro J, Navarro C (2018) Molecular detection of Canine Distemper Virus through the use of the Large Polymerase Gene. EASJALS 1(2): 39-43.
• 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 9(10): 2055-2064.