Molecular identification of Clostridium chauvoei from common filter paper / Molecular identification of Clostridium chauvoei from common filter paper

ABSTRACT

Background: Blackleg is an acute and often fatal infection in bovine caused by the bacterium Clostridium chauvoei. The absence of conclusive diagnosis of blackleg usually occurs due to absence of practical and economical methods to send samples to microbiology laboratory. The goal of this work was to verify the possibility of using ordinary fi lter paper as a practical and economically feasible method for collecting, storing and shipping material to the laboratory to be used in a rapid and direct PCR approach to detect Clostridium chauvoei DNA.Materials, Methods & Results: The PCR technique for the diagnosis of blackleg from common fi lter paper was tested for specifi city, sensitivity and feasibility. To test the specifi city, the papers were impregnated with a suspension of the following microorganisms C. chauvoei, C. perfringens, C. septicum, Bacillus anthracis, Staphylococcus aureus, and Escherichia coli. To test the sensitivity different concentration of C. chauvoei (ATCC 10092) were pipetted on common fi lter paper. To both test, DNA extraction of impregnated ordinary fi lter paper and their respective controls followed the method previously described and tested under different storage times (0 h, 24 h, 72 h and a week later). To test the feasibility, 12 bovine livers were collected and tissues samples were impregnate on common fi lter paper with suspension of C. cha

RESUMO

Background: Blackleg is an acute and often fatal infection in bovine caused by the bacterium Clostridium chauvoei. The absence of conclusive diagnosis of blackleg usually occurs due to absence of practical and economical methods to send samples to microbiology laboratory. The goal of this work was to verify the possibility of using ordinary fi lter paper as a practical and economically feasible method for collecting, storing and shipping material to the laboratory to be used in a rapid and direct PCR approach to detect Clostridium chauvoei DNA.Materials, Methods & Results: The PCR technique for the diagnosis of blackleg from common fi lter paper was tested for specifi city, sensitivity and feasibility. To test the specifi city, the papers were impregnated with a suspension of the following microorganisms C. chauvoei, C. perfringens, C. septicum, Bacillus anthracis, Staphylococcus aureus, and Escherichia coli. To test the sensitivity different concentration of C. chauvoei (ATCC 10092) were pipetted on common fi lter paper. To both test, DNA extraction of impregnated ordinary fi lter paper and their respective controls followed the method previously described and tested under different storage times (0 h, 24 h, 72 h and a week later). To test the feasibility, 12 bovine livers were collected and tissues samples were impregnate on common fi lter paper with suspension of C. cha