Direct Detection and Identification of African Trypanosomes by Fluorescence In Situ Hybridization with Peptide Nucleic Acid Probes

ABSTRACT We have developed a rapid and easy to perform fluorescence in situ hybridization test that allows specific identification of trypanosomes from the subgenus Trypanozoon, using peptide nucleic acid probes. Probes were designed to target subgenus-specific sequences on the multiple-copy 18S rRNA, greatly facilitating the detection of a single trypanosome.

[1]  S. Magez,et al.  Comparative Analysis of Antibody Responses against HSP60, Invariant Surface Glycoprotein 70, and Variant Surface Glycoprotein Reveals a Complex Antigen-Specific Pattern of Immunoglobulin Isotype Switching during Infection byTrypanosoma brucei , 2000, Infection and Immunity.

[2]  J. Coull,et al.  PNA solubility enhancers , 1998 .

[3]  Poonpilas Hongmanee,et al.  Fluorescence In Situ Hybridization Assay Using Peptide Nucleic Acid Probes for Differentiation between Tuberculous and Nontuberculous Mycobacterium Species in Smears of Mycobacterium Cultures , 1999, Journal of Clinical Microbiology.

[4]  D. Godfrey,et al.  Isolation of salivarian trypanosomes from man and other mammals using DEAE-cellulose. , 1970, Experimental parasitology.

[5]  W. Gibson,et al.  Identification of Trypanosoma brucei gambiense by PCR amplification of variant surface glycoprotein genes. , 1993, Acta tropica.

[6]  W. Gibson.  Will the real Trypanosoma b. gambiense please stand up. , 1986, Parasitology today.

[7]  G. Simo,et al.  Diagnosis of human trypanosomiasis, due to Trypanosoma brucei gambiense in central Africa, by the polymerase chain reaction. , 2000, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[8]  P. Simarro,et al.  Detection of Trypanosoma brucei gambiense in sleeping sickness suspects by PCR amplification of expression‐site‐associated genes 6 and 7 , 1999, Tropical medicine & international health : TM & IH.

[9]  A. Broomer,et al.  Identification of Dekkera bruxellensis(Brettanomyces) from Wine by Fluorescence In Situ Hybridization Using Peptide Nucleic Acid Probes , 2001, Applied and Environmental Microbiology.

[10]  F. Kansiime,et al.  Parasitological detection of Trypanosoma brucei gambiense in serologically negative sleeping-sickness suspects from north-western Uganda. , 1998, Annals of tropical medicine and parasitology.

[11]  W. Gibson,et al.  Detection of Trypanosoma brucei gambiense, in serologically positive but aparasitaemic sleeping-sickness suspects in Cameroon, by PCR. , 1996, Annals of tropical medicine and parasitology.

[12]  L. Vanhamme,et al.  Genetic controls for the expression of surface antigens in African trypanosomes. , 1994, Annual review of microbiology.

[13]  N. Meirvenne.  Biological diagnosis of human African trypanosomiasis , 1999 .

[14]  Peter E. Nielsen,et al.  PNA hybridizes to complementary oligonucleotides obeying the Watson–Crick hydrogen-bonding rules , 1993, Nature.

[15]  J. Corliss,et al.  The Trypanosomes of Mammals , 1973 .

[16]  E. Matovu,et al.  Detection of trypanosomes in suspected sleeping sickness patients in Uganda using the polymerase chain reaction. , 2000, Bulletin of the World Health Organization.

[17]  S. Rigby,et al.  Rapid detection, identification, and enumeration of Escherichia coli by fluorescence in situ hybridization using an array scanner. , 2001, Journal of microbiological methods.

[18]  W. Gibson,et al.  Absence of the LiTat 1.3 (CATT antigen) gene in Trypanosoma brucei gambiense stocks from Cameroon. , 1992, Acta tropica.

[19]  V. Jamonneau,et al.  Use of polymerase chain reaction in human African trypanosomiasis stage determination and follow-up. , 1999, Bulletin of the World Health Organization.

[20]  N. Van Meirvenne,et al.  A card-agglutination test with stained trypanosomes (C.A.T.T.) for the serological diagnosis of T. B. gambiense trypanosomiasis. , 1978, Annales de la Societe belge de medecine tropicale.

[21]  G. Hide.  History of Sleeping Sickness in East Africa , 1999, Clinical Microbiology Reviews.