학술논문

Loop-mediated isothermal amplification-based detection of typhoid fever on an automated Genie II Mk2 system – A case-control-based approach.
Document Type
Article
Source
Acta Tropica. Feb2019, Vol. 190, p293-295. 3p.
Subject
*TYPHOID fever
*GENE amplification
*ETHYLENEDIAMINETETRAACETIC acid
*SALMONELLA enterica
*BLOOD sampling
Language
ISSN
0001-706X
Abstract
Highlights • Two LAMP protocols for the detection of Salmonella enterica subsp. enterica serovar Typhi were assessed. • Both protocols showed reliable results with incubated blood culture materials with better sensitivity of the new approach. • For EDTA blood samples, sensitivity was low for both protocols. • qPCR for Salmonella enterica ssp. enterica ser. Typhi showed similar sensitivity with EDTA blood samples as LAMP. • The newly described LAMP-approach showed acceptable specificity even from EDTA blood. Abstract Typhoid fever, caused by the bacterium Salmonella enterica subsp. enterica serovar Typhi, is an important cause of blood stream infections in the tropics, for which easy-to-apply molecular diagnostic approaches are desirable. The diagnostic performance of a newly introduced and a previously described loop-mediated isothermal amplification (LAMP) approach using different primer sets on a Genie II Mk2 device for the identification of Salmonella enterica ssp. enterica ser. Typhi was evaluated with well-characterized residual materials from the tropics in a case control-based approach. After in-vitro confirmation of binding characteristics of both LAMP primer sets with culture isolates (n = 112), sensitivity and specificity were 100% for the newly designed new LAMP primer set 1 with incubated blood culture materials, while specificity was reduced to 97.1% for primer set 2. For 170 EDTA blood samples, sensitivity and specificity were 10% and 98.3% for primer set 1 as well as 38.0% and 83.3% for primer set 2, respectively; qPCR from EDTA blood did not score much better with 10% sensitivity and 100% specificity. LAMP using a Genie II Mk2 device is suitable for the identification of Salmonella enterica spp. enterica ser. Typhi from incubated blood culture materials. Sensitivity and specificity were insufficient for diagnosis directly from EDTA blood samples but LAMP showed similar sensitivity as qPCR. [ABSTRACT FROM AUTHOR]