Use of Chelex-100 for the molecular diagnosis of five animal pathogens

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

Revista Fave Ciencias Veterinarias

Universidad del Litoral

Lugar: Santa Fe; Año: 2021 vol. 20 p. 26 - 30

1666-938X

Molecular tools have improved conventional diagnosis in veterinary. Acid nucleic extraction is a key step fordownstream applications. This work aimed to compare4 DNA extraction methods: Chelex-100 resinwith Whatman® cards (M2), phenol- chloroform (M3), or commercial kits (M4) From animal pathogenseconomically or zoonotically important, receiving little attention, to determine the mostsensitive and inexpensive one for diagnosis. DNAwas isolated from urine,tissue, semen, blood and intestinal mucous, from the bacteria Leptospira30interrogans serovar Pomona Pomona, Brucella melitensis, and Salmonella ser. Abortusequi, and the parasites Leishmania spp., and Eimeria spp.respectively. The protocols sensitive was assayed by Polymerase Chain Reaction (PCR). TheM1 showed similar sensitivity for Salmonella ser. Abortusequi, Leishmania spp., and Eimeria spp., being better for L. interrogans serovar Pomona Pomona and slightly lower for B. melitensis.For the first time, a simple and economic method was successfully employed for extracting DNA fromthese animal pathogens, especially importantin low-resource settings, contributing to the diagnosis of leptospirosis, brucellosis, leishmaniasis, and coccidiosis; as well; as well as to the molecular epidemiology of salmonellosis in stallion from semen.