1921
Volume 70, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Two specific and sensitive polymerase chain reaction (PCR) assays were developed to detect and quantitate , the agent of scrub typhus, using a portion of the 47-kD outer membrane protein antigen/ high temperature requirement A gene as the target. A selected 47-kD protein gene primer pair amplified a 118-basepair fragment from all 26 strains of evaluated, but it did not produce amplicons when 17 and 18 less-related bacterial nucleic acid extracts were tested. Similar agent specificity for the real-time PCR assay, which used the same primers and a 31-basepair fluorescent probe, was demonstrated. This sensitive and quantitative assay determination of the content of nucleic acid used a plasmid containing the entire 47-kD gene from the Kato strain as a standard. Enumeration of the copies of DNA extracted from infected tissues from mice and monkeys following experimental infection with showed 27-5,552 copies/μL of mouse blood, 14,448-86,012 copies/μL of mouse liver/spleen homogenate, and 3-21 copies/μL of monkey blood.

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2004-04-01
2017-11-24
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  • Received : 24 Sep 2003
  • Accepted : 14 Dec 2003

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