Specific DNA probe for the sensitive detection of Trypanosoma evansi.

Abstract

Trypanosoma evansi is the parasitic protozoon that causes "Surra", a wasting disease of domestic animals. Detection of T. evansi plays an important role in epidemiology and animal health. DNA probes were constructed from T. evansi genomic DNA and kinetoplast DNA for sensitive detection of the parasite in infected blood. A 6.5 kb DNA insert of pMUT ec6 plasmid derived from the genomic DNA of T. evansi Npl isolate, selected from 575 recombinant E. coli exhibited the strongest nucleic acid hybridization signal to the T. evansi DNA. Using as the DNA probe, pMUT ec6 could detect as little as 60 pg T. evansi DNA and it did not hybridize to the DNA of cattle, waterbuffalo and two related blood parasites. A simple detection procedure by spotting 10 microliters infected blood onto nylon membrane could sense as little as 1000 parasites. The kinetoplast DNA was cloned in E. coli and found to show a comparable sensitivity to that of the pMUT ec6. However, the kinetoplast DNA exhibited variation in copy number among parasite isolates thus pMUT ec6 should be the DNA probe of choice for sensitive detection of T. evansi.