JVDI
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Journal of Veterinary Diagnostic Investigation Vol. 21 Issue 3, 354-358
Copyright © 2009 by the American Association of Veterinary Laboratory Diagnosticians
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Harada, K.
Right arrow Articles by Takahashi, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Harada, K.
Right arrow Articles by Takahashi, T.

Brief Research Reports

Comparison of three DNA extraction methods for detection of Erysipelothrix rhusiopathiae in chicken blood by polymerase chain reaction

Kazuki Harada1, Mariko Uchiyama, Teruyuki Hoshi and Toshio Takahashi

Correspondence: 1Corresponding Author: Kazuki Harada, National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo 185-8511, Japan. harada{at}nval.go.jp

A previously reported Erysipelothrix-specific polymerase chain reaction (PCR) was used to detect Erysipelothrix bacteremia in chickens. The sensitivity of PCR using 3 DNA extraction methods (boiling method, commercial gene matrix, and DNA extractor kit) was compared by using a serial 10-fold dilution of a chicken isolate of Erysipelothrix rhusiopathiae strain in chicken blood. Of the techniques used, the DNA extractor kit, followed by PCR, provided the most sensitive method for the detection of the E. rhusiopathiae strain in chicken blood (approximately 100 CFU/0.1 ml of blood). Two E. rhusiopathiae infection experiments were then attempted. In a total of 10 inoculated chickens, bacteremia developed in 9 chickens, consisting of all 5 chickens used in the first trial (ranging from 5.1 x 101 to 2.0 x 103 CFU/0.1 ml of blood) and 4 of the 5 chickens used in the second trial (ranging from 1.0 x 100 to 3.3 x 102 CFU/0.1 ml of blood). In the second trial, the 3 detection techniques were applied to the chickens with bacteremia, and the organism could be detected by using the DNA extractor kit in blood specimens from the 3 chickens exhibiting bacteremia of ≥4.2 x 101 CFU/0.1 ml of blood. This observation suggests that most E. rhusiopathiae–infected chickens develop more critical bacteremia than the detectable level by PCR with the DNA extractor kit, and the PCR detection method can be used as a first-line screening of avian erysipelas.

Key Words: Avian erysipelas • bacteremia • Erysipelothrix rhusiopathiae • rapid diagnosis






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2009 by the American Association of Veterinary Laboratory Diagnosticians, Inc.