A MUTATION SCANNING APPROACH FOR THE IDENTIFICATION OF HOOKWORM SPECIES AND ANALYSIS OF POPULATION VARIATION

To overcome limitations in the morphological identification of differe nt developmental stages of hookworms to species, we have established a polymerase chain reaction-linked single strand conformation polymorph ism technique (PCR-SSCP) utilizing the internal transcribed spacers (I TS) of ribosomal (r)DNA. These spacers were specifically chosen becaus e they provide reliable species markers for strongylid nematodes. ITS spacers were amplified by PCR from DNA derived from individual parasit es of seven species of hookworm, then denatured and subjected to elect rophoresis in a mutation detection enhancement (MDE) (non-denaturing) gel matrix. PCR-SSCP analysis showed that the single-strand ITS patter ns produced allowed the unequivocal identification of all species. The method also allowed the direct display of sequence variation within s ome species where multiple individual worms were examined. These findi ngs demonstrate the usefulness of the SSCP approach for hookworm ident ification, the detection of population variation and the direct displa y of sequence variation in rDNA. (C) 1998 Elsevier Science B.V. All ri ghts reserved.