PMID: 2116112Jan 1, 1990Paper

Detection of Rickettsia tsutsugamushi by gene amplification using polymerase chain reaction techniques

Annals of the New York Academy of Sciences
D J KellyM Carl

Abstract

Scrub typhus is commonly undiagnosed in endemic areas due, in part, to dependence on retrospective serodiagnosis. Since the etiologic agent, R. tsutsugamushi, will not grow in cell-free systems, a rapid direct-agent detection system such as provided by polymerase chain reaction (PCR) methodology is needed. Genes coding for the variable 56-kDa antigen of R. tsutsugamushi were amplified through 35 cycles using 20-mer oligonucleotide primers and Taq polymerase. Amplification of 1-ng samples of DNA extracted from purified prototype R. tsutsugamushi Karp, Gilliam, and Kato strains was detected by direct visual inspection of the electrophoresed, ethidium bromide-stained, specific bands. Specificity of the PCR was shown when PCR amplification of various non-scrub typhus rickettsial DNAs was unsuccessful. R. tsutsugamushi DNA extracted from the blood of infected mice could be PCR amplified and the 1477-base pair product detected by either direct visualization or by specific hybridization with amplified non-radioactive digoxigenin-11-dUTP-labeled Karp 56-kDa DNA probe.

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Citations

Oct 1, 1992·International Journal of Dermatology·Y TaniguchiM Shimizu
Nov 4, 2011·The American Journal of Tropical Medicine and Hygiene·Sang-Won ParkMyoung-don Oh
Apr 14, 2015·Future Microbiology·Alison Luce-FedrowAllen L Richards
Apr 22, 2006·Diagnostic Microbiology and Infectious Disease·Jiradej ManosroiAranya Manosroi
Feb 2, 2018·Clinical Microbiology Reviews·Fabián E DíazAlexis M Kalergis
May 1, 1991·European Journal of Epidemiology·H H Winkler
Dec 19, 2015·PLoS Neglected Tropical Diseases·Daryl J KellyAllen L Richards
Apr 1, 1992·The Journal of Dermatology·Y KannoK Ando
Oct 31, 2020·Tropical Medicine and Infectious Disease·Daniel H ParisAllen L Richards
Jun 1, 1994·Journal of Clinical Microbiology·S H KeeW H Chang

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