Loop-mediated isothermal amplification with the Procedure for Ultra Rapid Extraction kit for the diagnosis of pneumocystis pneumonia

Shuichi Kawano, Takuya Maeda, Takefumi Suzuki, Tatsuhiro Abe, Kei Mikita, Yusuke Hamakawa, Takeshi Ono, Wataru Sonehara, Yasushi Miyahira, Akihiko Kawana

研究成果: Article査読

8 被引用数 (Scopus)

抄録

Loop-mediated isothermal amplification (LAMP) is an innovative molecular technique requiring only a heating device and isothermal conditions to amplify a specific target gene. The results of current microscopic diagnostic tools for pneumocystis pneumonia are not sufficiently consistent for detecting infection with a low-density of Pneumocystis jirovecii. Although polymerase chain reaction (PCR) is highly sensitive, it is not suitable for resource-limited facilities. LAMP is a potential diagnostic replacement for PCR in such settings but a critical disadvantage of DNA extraction was still remained. Therefore, we employed the Procedure for Ultra Rapid Extraction (PURE) kit, which uses a porous material, to isolate the DNA from clinical samples in a simple way in combination with previously reported LAMP procedure for diagnosing PCP. The detection limit of the PURE-LAMP method applied to artificial bronchoalveolar lavage fluid samples was 100 copies/tube, even with the use of massive blood-contaminated solutions. In addition, we concluded the diagnostic procedure within 1 h without the need for additional equipment. PURE-LAMP coupled with suitable primers for specific pathogens has good potential for diagnosing various infectious diseases.

本文言語English
ページ(範囲)224-226
ページ数3
ジャーナルJournal of Infection and Chemotherapy
21
3
DOI
出版ステータスPublished - 2014 7 28
外部発表はい

ASJC Scopus subject areas

  • 微生物学(医療)
  • 薬理学(医学)
  • 感染症

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