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Extraction of microsporidial DNA from modified trichrome-stained clinical slides and subsequent species identification using PCR sequencing

  • K. S. CHAN (a1) and T. H. KOH (a1)

Molecular techniques involving polymerase chain reaction (PCR) and sequencing provide a relatively simple and objective means of identifying microsporidia to species level. We modified previously described methods of DNA extraction and PCR conditions for identification of microsporidia from museum slides, clinical specimens and environmental samples and successfully identified Vittaforma corneae in 11 out of 13 cases of microsporidial infection from used trichrome-stained slides of corneal scrapings from HIV-negative patients with keratoconjunctivitis.

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M. Hyliš , J. Weiser , M. Obornik and J. Vavra (2005). DNA isolation from museum and type collection slides of microsporidia. Journal of Invertebrate Pathology 88, 257260.

J. Joseph , S. Sharma , S. I. Murthy , P. V. Krishna , P. Garg , R. Nutheti , J. Kenneth and D. Balasubramanian (2006). Microsporidial keratitis in India: 16S rRNA gene-based PCR assay for diagnosis and species identification of microsporidia in clinical samples. Investigative Ophthalmology Visual Science 47, 44684473.

R. Weber , D. A. Schwartz and P. Deplazes (1999). Laboratory diagnosis of microsporidiosis. In The Microsporidia and Microsporidiosis (ed. M. Wittner and L. M. Weiss ), pp. 315362. ASM Press, Washington, DC.

L. M. Weiss and C. R. Vossbrinck (1999). Molecular biology, molecular phylogeny and molecular diagnostic approaches to the microsporidia. In The Microsporidia and Microsporidiosis (ed. M. Wittner and L. M. Weiss ), pp. 129171. ASM Press, Washington, DC.

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  • ISSN: 0031-1820
  • EISSN: 1469-8161
  • URL: /core/journals/parasitology
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