Uso de Chelex-100 para el diagnóstico molecular de cinco patógenos de animales

TOMAZIC MARIELA LUJÁN; ARREGUI MATIAS; WATANABE OLIVIA; GRUNE LOFFLER SYLVIA; HAMER, MICAELA; ASCENCIO MARIANO; BRIHUEGA BIBIANA.; BUSTOS CARLA; SARAULLO VANINA; RODRIGUEZ ANABEL ELISA

Revista Fave Ciencias Veterinarias

Facultad de Ciencias Agrarias de Esperanza (Universidad Nacional del Litoral)

Lugar: Santa Fe; Año: 2021 vol. 20

1666-938X

Molecular tools have improved conventional veterinary diagnosis. Acid nucleic extraction is a key step for downstream applications. This work aimed to compare the DNA extraction method Chelex-100 resin (M1) with Whatman® cards (M2), phenol-chloroform (M3),or commercial kits (M4), and to determine the most sensitive and inexpensive one for its diagnosis of animal pathogens that, despite their economic or zoonotic relevance, receive little attention. DNA was isolated from urine, organs, semen, blood and intestinal mucous, from the bacteria Leptospira interrogansserovar Pomona Pomona (by M1 and M2), Brucella melitensis(by M1, M3 and M4), and Salmonellaser. Abortusequi (by M1 and M4), and the parasites Leishmaniaspp. (by M1, M3 and M4), and Eimeria spp. (byM1 and M3), respectively. The sensitivity of each method was assayed by Polymerase Chain Reaction (PCR). The M1 showed similar sensitivity for Salmonellaser. Abortusequi, Leishmaniaspp., and Eimeriaspp., being better for L. interrogansserovar Pomona Pomona and slightly lower for B. melitensis. For the first time, a simple and economic method was successfully employed for extracting DNA from these animal pathogens, especially important in low-resource settings, contributing to the diagnosis of leptospirosis, brucellosis, leishmaniasis, and coccidiosis; as well as to the molecular epidemiology of salmonellosis in stallion from semen samples.