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A Nested PCR Assay to Detect DNA in Sera for the Diagnosis of Deep‐Seated Trichosporonosis
Author(s) -
Sugita Takashi,
Nakajima Masamitsu,
Ikeda Reiko,
Niki Yoshihito,
Matsushima Toshiharu,
Shinoda Takako
Publication year - 2001
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2001.tb01282.x
Subject(s) - nested polymerase chain reaction , biology , polymerase chain reaction , latex fixation test , direct agglutination test , microbiology and biotechnology , virology , gene , serology , antibody , immunology , genetics
Deep‐seated trichosporonosis caused by Trichosporon asahii has a high mortality rate and a very poor prognosis. New species‐specific oligonucleotide primers for T. asahii were developed from a sequence analysis of rRNA genes that included the internal transcribed spacer regions. A nested PCR assay with specific primers was used to examine 11 serum samples from 7 patients, who were diagnosed with deep‐seated trichosporonosis histologically at autopsy. In addition, Trichosporon cell wall polysaccharide (PS) was detected by a latex agglutination (LA) test. Of 11 samples, seven had a positive LA test, and T. asahii DNA was also detected with the nested PCR assay. Of the four samples in which PS antigen was not detected, the nested PCR of two samples was positive. Our new nested PCR assay may be used as an adjunct to conventional methods for diagnosing T. asahii infection.