PMID- 1626862
OWN - NLM
STAT- MEDLINE
DCOM- 19920813
LR  - 20190616
IS  - 0077-8923 (Print)
IS  - 0077-8923 (Linking)
VI  - 653
DP  - 1992 Jun 16
TI  - Evaluation of a DNA-based probe for the detection of cattle experimentally 
      infected with Babesia bigemina.
PG  - 131-45
AB  - A digoxigenin-labeled probe was used for hybridization to various preparations of 
      Babesia bigemina-infected erythrocyte extracts. Dot blot hybridization and 
      immunological detection of DNA hybrids revealed that the probe was specific for 
      B. bigemina DNA because it did not hybridize to the DNA of B. bovis, a closely 
      related species. Studies of sensitivity showed that the probe would bind to as 
      little as 1 ng of B. bigemina DNA, but not to 1 microgram of the B. bovis DNA. 
      The probe reacted with equal intensity against seven B. bigemina isolates from 
      different geographic areas. The lowest percentage of B. bigemina-infected 
      erythrocytes detected was 0.001%, a level of parasitemia not usually detected 
      with the light microscope. Six intact, mixed-breed steers, approximately 3 years 
      old, were inoculated with a blood stabilate containing B. bigemina-infected 
      erythrocytes. Blood samples collected from day -1 to day 86 postinoculation (PI) 
      and prepared for DNA extraction were analyzed in a dot blot hybridization assay 
      using a nonradioactive DNA probe. Hybridization reaction (HR) signals were 
      compared to results obtained by light microscopy (LM) examination of 
      Giemsa-stained blood smears and to antibody titers of serum samples assayed with 
      the complement fixation test (CFT) and indirect fluorescent antibody test (IFAT). 
      Four of six inoculated steers became infected with B. bigemina as assessed by LM. 
      The parasitemias were low (less than 0.01-0.05) at day 10 PI. Only three steers 
      were serologically positive by CFT (titer 1:40-1:160) and IFAT (1:1280). All four 
      infected steers had positive HR signals in the dot blot assay. The HR signals 
      were observed from day 10 to day 77 PI and were usually correlated with the 
      presence of parasites in blood as observed by LM. The HR signals varied in 
      intensity for different blood samples from the experimental animals and with day 
      of blood sample collection. Although the signal intensity did not correlate with 
      the parasitemia level estimated by LM, the nucleic acid hybridization assay was 
      more sensitive than LM, CFT, or IFAT for the detection of B. bigemina-infected 
      cattle.
FAU - Figueroa, J V
AU  - Figueroa JV
AD  - Department of Veterinary Microbiology, College of Veterinary Medicine, University 
      of Missouri, Columbia 65211.
FAU - Chieves, L P
AU  - Chieves LP
FAU - Byers, P E
AU  - Byers PE
FAU - Frerichs, W M
AU  - Frerichs WM
FAU - Buening, G M
AU  - Buening GM
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Ann N Y Acad Sci
JT  - Annals of the New York Academy of Sciences
JID - 7506858
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Protozoan)
SB  - IM
MH  - Animals
MH  - Babesia/genetics/*isolation & purification
MH  - Babesiosis/*diagnosis
MH  - Cattle
MH  - Cattle Diseases/*diagnosis
MH  - Colorimetry
MH  - *DNA Probes
MH  - DNA, Protozoan/*analysis/isolation & purification
MH  - Evaluation Studies as Topic
MH  - Male
MH  - Nucleic Acid Hybridization
MH  - Sensitivity and Specificity
EDAT- 1992/06/16 00:00
MHDA- 1992/06/16 00:01
CRDT- 1992/06/16 00:00
PHST- 1992/06/16 00:00 [pubmed]
PHST- 1992/06/16 00:01 [medline]
PHST- 1992/06/16 00:00 [entrez]
AID - 10.1111/j.1749-6632.1992.tb19636.x [doi]
PST - ppublish
SO  - Ann N Y Acad Sci. 1992 Jun 16;653:131-45. doi: 
      10.1111/j.1749-6632.1992.tb19636.x.