Oligonucleotide-gold nanoparticle networks for detection of Cryptosporidium parvum heat shock protein 70 mRNA

J Clin Microbiol. 2009 Dec;47(12):4060-6. doi: 10.1128/JCM.00807-09. Epub 2009 Oct 14.

Abstract

We report on a novel strategy for the detection of mRNA targets derived from Cryptosporidium parvum oocysts by the use of oligonucleotide-gold nanoparticles. Gold nanoparticles are functionalized with oligonucleotides which are complementary to unique sequences present on the heat shock protein 70 (HSP70) DNA/RNA target. The results indicate that the presence of HPS70 targets of increasing complexity causes the formation of oligonucleotide-gold nanoparticle networks which can be visually monitored via a simple colorimetric readout measured by a total internal reflection imaging setup. Furthermore, the induced expression of HSP70 mRNA in Cryptosporidium parvum oocysts via a simple heat shock process provides nonenzymatic amplification such that the HSP70 mRNA derived from as few as 5 x 10(3) purified C. parvum oocysts was successfully detected. Taken together, these results support the use of oligonucleotide-gold nanoparticles for the molecular diagnosis of cryptosporidiosis, offering new opportunities for the further development of point-of-care diagnostic assays with low-cost, robust reagents and simple colorimetric detection.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cryptosporidiosis / diagnosis*
  • Cryptosporidiosis / parasitology
  • Cryptosporidium parvum / genetics
  • Cryptosporidium parvum / growth & development
  • Cryptosporidium parvum / metabolism*
  • DNA, Protozoan / genetics
  • DNA, Protozoan / metabolism
  • Gold* / chemistry
  • HSP70 Heat-Shock Proteins / chemistry
  • HSP70 Heat-Shock Proteins / genetics
  • HSP70 Heat-Shock Proteins / metabolism*
  • Humans
  • Nanoparticles* / chemistry
  • Oligonucleotides* / chemistry
  • Oligonucleotides* / genetics
  • Oocysts / growth & development
  • Oocysts / metabolism
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sensitivity and Specificity

Substances

  • DNA, Protozoan
  • HSP70 Heat-Shock Proteins
  • Oligonucleotides
  • RNA, Messenger
  • Gold