Development and evaluation of an extra chromosomal DNA-based PCR test for diagnosing bovine babesiosis.
Subclinical infections of bovine babesiosis, caused primarily by Babesia bigemina or Babesia bovis are a challenge to current diagnostic methods. In this study, the development and evaluation of a PCR test for sensitive and specific detection of B. bigemina or B. bovis is described. The target selected for amplification is part of the apocytochrome b gene, conserved in both Babesia spp. and located on the linear approximately 6.0 kb extra chromosomal DNA. The test was evaluated to detect the parasites over a period of 5 (B. bigemina) and 10 months (B. bovis) post infection in experimentally infected cattle. Analysis of DNA extracted from blood samples drawn from the experimental cattle in a blind study revealed an overall sensitivity of 85 and 64% for B. bovis and B. bigemina respectively, while the specificity was 97% for B. bovis and 91% for B. bigemina. The test results were compared with the recently developed ribosomal DNA-based polymerase chain reaction (PCR) test and to the complement fixation test for both Babesia spp. The extra chromosomal DNA-based test was 20% more sensitive than that of ribosomal DNA-based tests. This test may be a more desirable alternative to the currently used, complement fixation test.
Salem
G H
GH
College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
Liu
X
X
Johnsrude
J D
JD
Dame
J B
JB
Roman Reddy
G
G
D000818:Animals; D001059:Apoproteins / Q000235:genetics; D001403:Babesia / Q000235:genetics / Q000302:isolation & purification*; D016793:Babesia bovis / Q000235:genetics / Q000302:isolation & purification*; D001404:Babesiosis / Q000175:diagnosis* / Q000469:parasitology; D002353:Carrier State / Q000175:diagnosis / Q000662:veterinary*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology; D003168:Complement Fixation Tests; D003573:Cytochrome b Group / Q000235:genetics; D045303:Cytochromes b; D016054:DNA, Protozoan / Q000097:blood / Q000235:genetics*; D004275:DNA, Ribosomal / Q000097:blood / Q000235:genetics; D005111:Extrachromosomal Inheritance / Q000235:genetics*; D015345:Oligonucleotide Probes; D018512:Parasitemia / Q000469:parasitology / Q000662:veterinary*; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D016037:Single-Blind Method; D013045:Species Specificity
D001059:Apoproteins; D003573:Cytochrome b Group; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D015345:Oligonucleotide Probes; D015800:Protozoan Proteins; D045303:Cytochromes b