BioTechniques (Jan 2001)

Hybridization Screening of Very Short PCR Products for Paleoepidemiological Studies of Chagas’ Disease

  • M. Madden,
  • W.L. Salo,
  • J. Streitz,
  • A.C. Aufderheide,
  • G. Fornaciari,
  • C. Jaramillo,
  • G.A. Vallejo,
  • R. Yockteng,
  • B. Arriaza,
  • F. Cárdenas-Arroyo,
  • F. Guhl

DOI
https://doi.org/10.2144/01301st07
Journal volume & issue
Vol. 30, no. 1
pp. 102 – 109

Abstract

Read online

Single strands of very short PCR products can be covalently immobilized to a slide and then easily detected by probe hybridization. In this work, the PCR product was a 70-nucleotide segment of ancient DNA, representing a portion of repeat minicircle DNA from the kinetoplast of Trypanosoma cruzi, the infectious agent of Chagas’ disease (American Trypanosomiasis). The target segment was initially established to be present in soft tissue samples taken from four “naturally” mummified Andean bodies using PCR followed by cloning and sequencing. Hybridization screening of the covalently immobilized PCR products positively identified products from 25 of 27 specimens of different tissues from these four mummies. The method appears to be ideal for the purpose of screening a large number of specimens when the target PCR product is very short.