Skip to main content

Sleeping sickness in Zaire: a nested polymerase chain reaction improves the identification of Trypanosoma ( Trypanozoon ) brucei gambiense by specific kinetoplast DNA probes

Buy Article:

$51.00 plus tax (Refund Policy)

Abstract:

Blood samples collected in the sleeping sickness focus of Boma, Zaire, from human patients and domestic animals were analysed by polymerase chain reaction (PCR) for the presence of trypanosome DNA. The comparison of PCR and miniature anion exchange centrifugation technique (m‐AECT) results clearly shows that in domestic animals mixed infections (Trypanozoon/Trypanosoma [Nannomonas] congolense) are more frequently diagnosed by PCR than by m‐AECT. Trypanozoon positive blood samples were further analysed for Trypanosoma (Trypanozoon) brucei gambiense. For that purpose amplified minicircle kinetoplast DNA (minicircle kDNA) was differentiated in gambiense and non‐gambiense by hybridization with DNA probes. To analyse blood samples, especially those with low parasite numbers, the amplification step had to be improved by a nested PCR. Subsequent hybridiz‐ation was done with kDNA probes generated by PCR from blood samples which had been obtained from a human patient infected with T. (T.) b. gambiense and a pig infected with Trypanozoon. The hybridization results clearly show that at least two genotypes of Trypanozoon parasites occur in the sleeping sickness focus of Boma, Bas‐Zaire. One obviously corresponds to T. (T.) b. gambiense and was present in humans and two domestic animals (pig, dog). The other genotype seems to be associated with T. (T.) b. brucei and could be detected only in the blood of domestic animals. This is the first time that field samples could be analysed by a technique which facilitates the molecular identification of T. (T.) b. gambiense without prior cloning, propagation, and/or isolation of the parasites. Therefore, this technique seems to be a promising tool to elucidate the significance of the animal reservoir for the epidemiology of the gambiense sleeping sickness in Africa.

Keywords: PCR; Trypanosoma brucei gambiense; Trypanosoma brucei rhodesiense; Trypanosoma congolense; hybridization; m‐AECT

Document Type: Original Article

DOI: http://dx.doi.org/10.1046/j.1365-3156.1996.d01-11.x

Affiliations: Institute of Parasitology and Tropical Veterinary Medicine, Free University Berlin, Germany

Publication date: February 1, 1996

bsc/tmih/1996/00000001/00000001/art00008
dcterms_title,dcterms_description,pub_keyword
6
5
20
40
5

Access Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content
Cookie Policy
X
Cookie Policy
ingentaconnect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more