Sequence conservation of repeat 3 region of the gene coding for the 15 kDa polyprotein within human and simian Loa loa.
The human and simian strains of Loa loa microfilariae are morphologically identical even though their periodicities vary. When using primate models (Mandrillus sphinx) of human loaisis for vaccination trials, the absence of any ongoing simian L. loa infection must be demonstrated. Nested primers derived from a human strain of L. loa (targeted on the repeat 3 region of the gene encoding the 15 kDa polyprotein; 15r3) amplified at 366 bp sequence from simian L. loa genomic DNA and blood lysates from mandrills infected with simian L. loa. This nested-PCR assay has been tested on 12 amicrofilaremic (AMF) mandrills (without filarial microfilariae) and was positive in four mandrills. The nested-PCR product derived from simian L. loa genomic DNA and from three of four AMF mandrills has been sequenced. No difference was observed between the four sequences, which, in addition, were 99.18% identical to the 15r3 of human L. loa. Therefore, the 15r3 sequence is conserved within human and simian L. loa. These results suggest that the four PCR-positive mandrills without circulating microfilariae had occult simian L. loa infections. The study demonstrates the ability of a nested-PCR assay to identify animals naturally infected with simian L. loa.
Toure
F S
FS
Centre International de Recherches Médicales de Franceville, Gabon. ftoure@cirmf.sci.ga
Leroy
E M
EM
Mavoungou
E
E
Egwang
T G
TG
D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics*; D001483:Base Sequence; D017124:Conserved Sequence / Q000235:genetics*; D018509:DNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D015801:Helminth Proteins / Q000235:genetics*; D006801:Humans; D008117:Loa / Q000235:genetics* / Q000302:isolation & purification; D008118:Loiasis / Q000469:parasitology* / Q000662:veterinary; D008969:Molecular Sequence Data; D008992:Monkey Diseases / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D017422:Sequence Analysis, DNA
C095839:15kDa ladder antigen, Loa loa; D000947:Antigens, Helminth; D018509:DNA, Helminth; D015801:Helminth Proteins