Rapid diagnosis of leishmaniasis by fluorogenic polymerase chain reaction

被引：83
|
作者：
Wortmann, G
Sweeney, C
Houng, HS
Aronson, N
Stiteler, J
Jackson, J
Ockenhouse, C
机构：
[1] Walter Reed Army Med Ctr, Washington, DC 20307 USA
[2] Walter Reed Army Inst Res, Silver Spring, MD 20910 USA
[3] Uniformed Serv Univ Hlth Sci, Div Infect Dis, Bethesda, MD 20814 USA
来源：
AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE | 2001年 / 65卷 / 05期
关键词：
D O I：
10.4269/ajtmh.2001.65.583
中图分类号：
R1 [预防医学、卫生学];
学科分类号：
1004 ; 120402 ;
摘要：
A fluorescent DNA probe (LEIS.P1) specific for a conserved region of the small-subunit ribosomal RNA gene of Leishmania and a pair of flanking primers (LEIS.U1 and LEIS.L1) were designed for use in a fluorogenic polymerase chain reaction. Optimal assay conditions with zero background were established to detect low levels of Leishmania from clinical samples. By use of this assay, we amplified DNA from 27 strains of cultured Leishmania (both Old and New World strains) and selectively amplified Leishmania DNA from 12 paraffin-embedded human biopsy samples and 3 fresh human skin biopsy specimens. For the fresh human tissue biopsies, the turnaround time from biopsy to test result was < 24 hr. No amplification was detected in negative control samples (including the kinetoplastid protozoa Trypanosoma rangelli and Crithidia fasiculata). This assay provides a specific and rapid diagnostic modality to detect infection with Leishmania.
引用
收藏
页码：583 / 587
页数：5
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