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

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)224-226
Number of pages3
JournalJournal of Infection and Chemotherapy
Volume21
Issue number3
DOIs
Publication statusPublished - 2014 Jul 28
Externally publishedYes

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Keywords

  • Loop-mediated isothermal amplification
  • Pneumocystis jirovecii
  • Pneumocystis pneumonia
  • Polymerase chain reaction
  • Procedure for Ultra Rapid Extraction

ASJC Scopus subject areas

  • Microbiology (medical)
  • Pharmacology (medical)
  • Infectious Diseases
  • Medicine(all)

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