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Title: MULTIPLEX PCR FOR THE IDENTIFICATION OF ARCOBACTER AND DIFFERENTIATION OF ARCOBACTER BUTZLERI FROM OTHER ARCOBACTERS

Author
item HARMON, K - IOWA STATE UNIVERSITY
item WESLEY, IRENE

Submitted to: Veterinary Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/25/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary: Arcobacter spp. are a newly recognized group of bacteria. They resemble and are closely related to Campylobacter jejuni, which is a major cause of human foodborne illness. Arcobacter butzleri is present in clinically healthy livestock. It has also been implicated in cases of enteritis in humans and animals as well as abortion in livestock. The Arcobacter bacteria are difficult to grow in the laboratory, which complicates their detection in animal tissues and in clinical specimens. We developed a DNA test based on the polymerase chain reaction (PCR) assay to simultaneously detect Arcobacter and to identify A. butzleri in a single test. The assay is accurate, simple to perform, and the results are easily interpreted. The entire assay requires less than eight hours to complete, compared to several days or weeks for current methods. This test will assist in determining the prevalence of Arcobacter in livestock and in meats. Information gained from use of this test in surveys will benefit governmen agencies to check for bacteria in food.

Technical Abstract: The genus Arcobacter encompasses gram-negative, aerotolerant, spiral-shaped bacteria. Arcobacter spp. are difficult to grow in the laboratory and phenotypic traits for species differentiation are limited. Arcobacter butzleri has been isolated from livestock, drinking water, poultry, and ground pork as well as from cases of human enteritis. Herein we describe a amultiplex polymerase chain reaction (PCR) assay to identify Arcobacter isolates and to distinguish A. butzleri from other arcobacters. The test uses two primer sets. Set I targets a section of the 16S rRNA genes of Arcobacter spp. Set II amplifies a portion of the 23S rRNA genes unique to A. butzleri. Specificity of the primer sets was evaluated using ATCC reference strains. Upon PCR amplification, all of the Arcobacter isolates yielded a 1223 bp product. Arcobacter butzleri ATCC 49616 exhibited both a 1223 bp and a 686 bp product. Arcobacter cryaerophilus ATCC 43159, which was identified by slot blot as A. butzleri, also yielded the two amplicons typical of A. butzleri. No PCR product was observed for closely related ATCC strains (n=37). We next analyzed by multiplex PCR field strains of Arcobacter spp. (n=108) which had been previously characterized to the species level by either DNA-DNA hybridization, dot blot hybridization, ribotyping, or by serology. Speciation by multiplex PCR agreed with results obtained by the other methods. The multiplex PCR assay is specific, rapid, and easy to interpret and thus will aid in elucidating the prevalence, epidemiology, and zoonotic potential of Arcobacter.