Evaluation of the Brucella abortus species-specific polymerase chain reaction assay, an improved version of the Brucella AMOS polymerase chain reaction assay for cattle.

نویسندگان

  • Betsy J Bricker
  • Darla R Ewalt
  • Steven C Olsen
  • Allen E Jensen
چکیده

In a blind test, 344 samples representing 80 bacterial isolates were analyzed by the Brucella abortus species-specific polymerase chain reaction (BaSS PCR) assay for the identification and discrimination of B. abortus field strains (wild-type biovars 1, 2, and 4) from 1) B. abortus vaccine strains, 2) other Brucella species, and 3) non-Brucella bacteria. Identical samples were tested in 2 laboratories. Half the samples were fully viable, and half were bacteria that had been killed by methanol fixation. The results in 1 laboratory correctly identified 100% of the samples, resulting in a predictive value of 100% for all categories and 100% sensitivity and specificity under the prescribed conditions. The second laboratory misidentified 31 samples, resulting in a range of 66.7-100% sensitivity, 93.2-99.7% specificity, and 77.3-98.2% predictive values depending on the category. There was no significant difference in viable versus fixed bacteria for either laboratory. Subsequent review of the protocol indicated that contamination was the likely cause of 26 of the 31 erroneous identifications. The results show that the BaSS PCR assay has the potential to be a very reliable screening tool for B. abortus identification. However, the data also provide a cautionary reminder of the importance of preventing contamination in diagnostic PCR.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Development of New Modified Simple Polymerase Chain Reaction and Real-time Polymerase Chain Reaction for the Identification of Iranian Brucella abortus Strains

Brucellosis is primarily a worldwide zoonotic disease caused by Brucella species. The genus Brucella contains highly infectious species that are classified as biological threat agents. In this regard, the identification of Brucella can be a time-consuming and labor-intensive process posing a real risk of laboratory-acquired infection to the laboratory staff. This stud...

متن کامل

Development of a multiplex polymerase chain reaction assay for differentiation of field strain isolates and vaccine strains S19 and RB51 of Brucella in Iran

Bovine brucellosis is a zoonotic disease distributed worldwide and characterized by abortion and reducedfertility in cows. Since brucellosis eradication programme in Iran uses vaccination, test, slaughter andquarantine as control measures, it is essential to distinguish vaccine strains from strains that cause infectionsamong vaccinated cattle herds. We developed and evaluated a multiplex polyme...

متن کامل

Detection of Brucella abortus in Chiredzi district in Zimbabwe.

Brucellosis is an endemic disease in Zimbabwe caused by the genus Brucella. Brucella seroprevalence was recently reported to be high in the wildlife-livestock interface in the Chiredzi district and the neighbouring Gonarezhou National Park (GNP) in Zimbabwe, and higher amongst communal cattle with an abortion history and access to grazing in GNP than amongst communal cattle with no aborti...

متن کامل

Comparative efficacy of serological diagnostic methods and evaluation of polymerase chain reaction for diagnosis of bovine brucellosis

The aim of present study was to compare the diagnostic performance of the different Brucella abortus antigen based serological and molecular tests such as Rose Bengal Plate Test (RBPT), indirect enzyme linked immunosorbent assay (I-ELISA) and polymerase chain reaction (PCR). Out of 89 samples, 44 were positive by I-ELISA, 18 by RBPT and 21 by PCR. Substantial agreement was observed between PCR ...

متن کامل

A sensitive & specific multiplex PCR assay for simultaneous detection of Bacillus anthracis, Yersinia pestis, Burkholderia pseudomallei & Brucella species

BACKGROUND & OBJECTIVES Bacillus anthracis, Yersinia pestis, Burkholderia pseudomallei and Brucella species are potential biowarfare agents. Classical bacteriological methods for their identification are cumbersome, time consuming and of potential risk to the handler. METHODS We describe a sensitive and specific multiplex polymerase chain reaction (mPCR) assay involving novel primers sets for...

متن کامل

ذخیره 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 نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

دوره 15 4  شماره 

صفحات  -

تاریخ انتشار 2003