Parasitology

Research Article

Detection of Trypanosoma congolense and Trypanosoma brucei subspecies by DNA amplification using the polymerase chain reaction

D. R. Mosera1, G. A. Cooka1 p1, Diane E. Ochsa1, Cheryl P. Baileya1, Melissa R. McKanea1 and J. E. Donelsona1

a1 Department of Biochemistry, and Diabetes and Endocrinology Research Center, University of Iowa, Iowa City, IA 52242

Abstract

The nuclear DNA of Trypanosoma congolense contains a family of highly conserved 369 base pair (bp) repeats. The sequences of three cloned copies of these repeats were determined. An unrelated family of 177 bp repeats has previously been shown to occur in the nuclear DNA of Trypanosoma brucei brucei (Sloof et al. 1983a). Oligonucleotides were synthesized which prime the specific amplification of each of these repetitive DNAs by the polymerase chain reaction (PCR). Amplification of 10% of the DNA in a single parasite of T. congolense or T. brucei spp. produced sufficient amplified product to be visible as a band in an agarose gel stained with ethidium bromide. This level of detection, which does not depend on the use of radioactivity, is about 100 times more sensitive than previous detection methods based on radioactive DNA probes. The oligonucleotides did not prime the amplification of DNA sequences in other trypanosome species nor in Leishmania, mouse or human DNAs. Amplification of DNA from the blood of animals infected with T. congolense and/or T. brucei spp. permitted the identification of parasite levels far below that detectable by microscopic inspection. Since PCR amplification can be conducted on a large number of samples simultaneously, it is ideally suited for large-scale studies on the prevalence of African trypanosomes in both mammalian blood and insect vectors.

(Accepted February 08 1989)

Key words

  • Trypanosoma congolense;
  • Trypanosoma brucei;
  • DNA amplification;
  • polymerase chain reaction.

Correspondence:

p1 Department of Internal Medicine, University of Iowa, Iowa City, Iowa 52242, USA.

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