Use of Multiplex Real-Time PCR To Diagnose Scrub Typhus


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Comparison of conventional, nested, and real-time quantitative PCR for diagnosis of scrub typhus.

Orientia tsutsugamushi is the causative agent of scrub typhus. For the diagnosis of scrub typhus, we investigated the performances of conventional PCR (C-PCR), nested PCR (N-PCR), and real-time quantitative PCR (Q-PCR) targeting the O. tsutsugamushi-specific 47-kDa gene. To compare the detection sensitivities of the three techniques, we used two template systems that used plasmid DNA (plasmid d...

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Real-time multiplex PCR assays.

The ability to multiplex PCR by probe color and melting temperature (T(m)) greatly expands the power of real-time analysis. Simple hybridization probes with only a single fluorescent dye can be used for quantification and allele typing. Different probes are labeled with dyes that have unique emission spectra. Spectral data are collected with discrete optics or dispersed onto an array for detect...

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Usefulness of eschar PCR for diagnosis of scrub typhus.

We report here on the case of a child who was infected with scrub typhus, and we made the diagnosis according to the serology and by performing PCRs on the child's eschar. The patient was treated with azithromycin, and he did not experience any complications. Performing nested PCR on the eschar might be both a rapid diagnostic test for scrub typhus in the early acute stage and a differential te...

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Scrub typhus is an important emerging tropical infection that can have severe manifestations in children leading to intensive care admission. It is caused by Orientia tsutsugamushi, a gram negative bacterium transmitted to humans by the bite of larval trombiculid mites. The organism is endemic in south-east Asia and the disease is reported from virtually all parts of India. The hallmark of this...

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Key words: herpes simplex virus, real-time PCR, multiplex real-time PCR assay

要 旨 単 純 ヘ ル ペ ス ウ イ ル ス1型(HSV-1)お よ び2型(HSV-2)の 遺 伝 子 を 同時 に検 出 す るmultiplex real-time PCR法 を 開発 し た. HSV-1お よびHSV-2のgpD領 域 を標 的 と し,プ ラ イ マ ー お よ びTaqMan蛍 光 プ ロ ー ブ を 独 自 に 設 計 し た.検 出感 度 は15copies/tubeで,HSV-1お よ びHSV-2と も15か ら1.5×106 copies/tubeの 範 囲 で 定 量 が 可 能 で あ っ た.real-time PCR法 はPCR法 よ り も103倍 感 度 が 高 く,他 の ヘ ルペ ス属(CMV, EBV, HHV6, HHV-7お よ びVZV) DNAは 検 出 され なか っ た. 以 上 の結 果 か ら,今 回 開 発...

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عنوان ژورنال: Journal of Clinical Microbiology

سال: 2017

ISSN: 0095-1137,1098-660X

DOI: 10.1128/jcm.02181-16