Detection of cryptococcus neoformans gene using pcr.

Ken ichi Tanaka, Shigeru Kohno, Kohtaro Mitutake, Haruko Miyazaki, Shigefumi Maesaki, Tetsuhiro Noda, Takashige Miyazaki, Mitsuo Kaku, Hironobu Koga, Kouhei Hara

Research output: Contribution to journalArticle

Abstract

We studied detection of the Cryptococcus neoformans gene using PCR method. The primer was designed from 20 mer of Cryptococcus neoformans URA 5 gene. Conditions of the PCR method were denaturing at 94°C for 60sec, annealing at 63°C for 90 sec, extension 72°C for 60 sec, and this was repeated for 36 cycles. DNA extraction was conducted by the glass bead method, the enzyme method or a combination of the two and the glass bead method showed the highest extraction rate in experiments. The series of experiments revealed the high specificity of the adapted PCR method. Sensitivity in nested PCR was 100pg in quantity of DNA and 1×103 in diluted solution.

Original languageEnglish
Pages (from-to)141-143
Number of pages3
JournalJapanese Journal of Medical Mycology
Volume35
Issue number2
DOIs
Publication statusPublished - 1994 Jan 1

Keywords

  • cryptococcosis
  • polymelase chain reaction

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

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  • Cite this

    Tanaka, K. I., Kohno, S., Mitutake, K., Miyazaki, H., Maesaki, S., Noda, T., Miyazaki, T., Kaku, M., Koga, H., & Hara, K. (1994). Detection of cryptococcus neoformans gene using pcr. Japanese Journal of Medical Mycology, 35(2), 141-143. https://doi.org/10.3314/jjmm.35.141