Use of an enrichment broth cultivation-PCR combination assay for rapid diagnosis of swine erysipelas.
نویسندگان
چکیده
We have previously described the creation by Tn916 mutagenesis of avirulent transposition mutants from a highly virulent strain of Erysipelothrix rhusiopathiae, the causative agent of swine erysipelas. In this study, we cloned a 2.2-kb DNA fragment which flanked the Tn916 insertion in an avirulent mutant (strain 33H6) and evaluated the possibility that this region could be used for the specific detection of E. rhusiopathiae. According to the sequences of this region, oligonucleotide primers were designed to amplify a 937-bp fragment of the E. rhusiopathiae chromosome by PCR. The specificity of the PCR was investigated by analyzing 64 strains of Erysipelothrix species and 27 strains of other genera different from Erysipelothrix. A 937-bp DNA fragment could be amplified from all E. rhusiopathiae strains tested, and no amplification was observed by using DNAs from the other species tested. To make a rapid and definite diagnosis of swine erysipelas in slaughterhouses, we developed an enrichment broth cultivation-PCR combination assay, which used a commercially available DNA extraction kit, to identify E. rhusiopathiae in the specimens from swine with arthritis. After samples were enriched in selective broth culture, detection of E. rhusiopathiae was tested by either conventional methods or the PCR. Of 102 samples tested, 15 samples were positive by conventional methods and 12 of the 15 samples were positive by the PCR. The detection limit of the PCR was 10(3) CFU per reaction mixture for the PCR-positive samples. These results indicate that this PCR technique could be used as a first-line screening technique for the specific detection of E. rhusiopathiae in specimens.
منابع مشابه
Swine Erysipelas Combination Assay for Rapid Diagnosis of Use of an Enrichment Broth Cultivation-PCR
متن کامل
Comparison of cultivation and PCR-hybridization for detection of Salmonella in porcine fecal and water samples.
A total of 150 fecal and water samples from four swine farms were tested for the presence of Salmonella enterica using different enrichment techniques as follows: (i) 92 fecal samples from nursery and farrowing barns at three swine farms were preenriched overnight in tryptic soy broth (TSB) at 37 degrees C followed by overnight enrichment in Rappaport-Vassiliadis 10 broth (RV10) at 42 degrees C...
متن کاملRapid and accurate diagnosis of Foot-and-mouth disease virus by Real-time PCR in field samples
During 2010-2011, Real-time PCR procedure was used to detecting FMDV RNA on 147 epithelium samples from the field. In this survey, for Real-time PCR from 3D gene segment as conserve region selected for tracking all of seven serotypes FMDV. The assay detected the viral RNA in all serotypes of FMDV. The rRT-PCR specifically detected FMD virus in sample with greater sensitivity than our convention...
متن کاملRapid detection of vanA and vanB genes directly from clinical specimens and enrichment broths by real-time multiplex PCR assay.
A real-time PCR assay previously developed for use on the Roche LightCycler platform was investigated as an alternative to culture for the direct detection of vancomycin-resistant enterococci (VRE) in clinical specimens. PCR primers and fluorescence resonance energy transfer hybridization probes specific for the vanA and vanB genes were combined in a multiplex real-time PCR assay performed dire...
متن کاملComparison of Dna-extraction Methods and Selective Enrichment Broths on the Detection of Salmonella Typhimurium in Swine Feces by Polymerase Chain Reaction (pcr)
The aim of this study was to compare different DNA-extraction methods and selective enrichment broths for their effectiveness to detect Salmonella Typhimurium in artificially inoculated swine feces samples (100 CFU/ g) by polymerase chain reaction. After enrichment in Rappaport-Vassiliadis, selenite cystine or MüllerKauffmann tetrathionate, aliquots were used for DNA extraction by three differe...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of clinical microbiology
دوره 36 1 شماره
صفحات -
تاریخ انتشار 1998