Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen

Background Typhoid fever caused by Salmonella typhi is a common acute infection of the reticuloendothelial system, intestinal lymphoid tissue, and gall bladder. Detection of Salmonella spp. is still based on cultures and serological methods.Widal test is one of the serological tests that is still...

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Autores principales:	Oktania Sandra Puspita, Andi Yasmon, Beti Ernawati Dewi
Formato:	article
Lenguaje:	EN
Publicado:	Diponegoro University 2020
Materias:	salmonella enterica subsp.enterica typhoid fever ssan gene real time pcr Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/8165b8bc890e4f24bc4dc4afd69c110c
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spelling	oai:doaj.org-article:8165b8bc890e4f24bc4dc4afd69c110c2021-11-05T16:47:42ZOptimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen2503-217810.14710/jbtr.v6i2.7120https://doaj.org/article/8165b8bc890e4f24bc4dc4afd69c110c2020-08-01T00:00:00Zhttps://ejournal2.undip.ac.id/index.php/jbtr/article/view/7120https://doaj.org/toc/2503-2178Background Typhoid fever caused by Salmonella typhi is a common acute infection of the reticuloendothelial system, intestinal lymphoid tissue, and gall bladder. Detection of Salmonella spp. is still based on cultures and serological methods.Widal test is one of the serological tests that is still widely used, especially in developing countries including Indonesia.Widal tests have low sensitivity and specificity. They often produce false positive or false negative results. Objective The aim of this study were i) real time PCR optimization to develop a Salmonella enterica detection system. ii) molecular detection of new target gene (ssaN gene) from blood specimens in typhoid fever patients. Methods An experimental laboratory study was performed from March to October 2016. Extraction of Salmonella typhi DNA is used as templates for the optimization of real time PCR reaction.The blood sample was from patients suspected with typhoid fever obtained from the Menteng Sub-district Health Center according to the inclusion criteria. Results Specificity test of real time PCR showed that the primers and probes used are not cross-react against other microorganisms. Sensitivity test obtained minimal detection is at least 10 cfu/ml of blood specimen. In blood clinical specimens, real time PCR could detect 19 (38%) positive samples of 50 blood specimen from suspected typhoid fever patients. Eleven samples with negative Widal serology gives positive results in real time PCR. Conclusion Real time PCR used in this study can increase the level of rate of positive testing by 22% of the total specimens. Keywords : Salmonella enterica subsp.enterica, typhoid fever, ssaN gene, real time PCROktania Sandra PuspitaAndi YasmonBeti Ernawati DewiDiponegoro Universityarticlesalmonella enterica subsp.entericatyphoid feverssan genereal time pcrMedicine (General)R5-920ENJournal of Biomedicine and Translational Research, Vol 6, Iss 2, Pp 41-47 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	salmonella enterica subsp.enterica typhoid fever ssan gene real time pcr Medicine (General) R5-920
spellingShingle	salmonella enterica subsp.enterica typhoid fever ssan gene real time pcr Medicine (General) R5-920 Oktania Sandra Puspita Andi Yasmon Beti Ernawati Dewi Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen
description	Background Typhoid fever caused by Salmonella typhi is a common acute infection of the reticuloendothelial system, intestinal lymphoid tissue, and gall bladder. Detection of Salmonella spp. is still based on cultures and serological methods.Widal test is one of the serological tests that is still widely used, especially in developing countries including Indonesia.Widal tests have low sensitivity and specificity. They often produce false positive or false negative results. Objective The aim of this study were i) real time PCR optimization to develop a Salmonella enterica detection system. ii) molecular detection of new target gene (ssaN gene) from blood specimens in typhoid fever patients. Methods An experimental laboratory study was performed from March to October 2016. Extraction of Salmonella typhi DNA is used as templates for the optimization of real time PCR reaction.The blood sample was from patients suspected with typhoid fever obtained from the Menteng Sub-district Health Center according to the inclusion criteria. Results Specificity test of real time PCR showed that the primers and probes used are not cross-react against other microorganisms. Sensitivity test obtained minimal detection is at least 10 cfu/ml of blood specimen. In blood clinical specimens, real time PCR could detect 19 (38%) positive samples of 50 blood specimen from suspected typhoid fever patients. Eleven samples with negative Widal serology gives positive results in real time PCR. Conclusion Real time PCR used in this study can increase the level of rate of positive testing by 22% of the total specimens. Keywords : Salmonella enterica subsp.enterica, typhoid fever, ssaN gene, real time PCR
format	article
author	Oktania Sandra Puspita Andi Yasmon Beti Ernawati Dewi
author_facet	Oktania Sandra Puspita Andi Yasmon Beti Ernawati Dewi
author_sort	Oktania Sandra Puspita
title	Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen
title_short	Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen
title_full	Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen
title_fullStr	Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen
title_full_unstemmed	Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen
title_sort	optimizing real-time pcr methods for detection of ssan gene salmonella enterica subsp.enterica in the blood specimen
publisher	Diponegoro University
publishDate	2020
url	https://doaj.org/article/8165b8bc890e4f24bc4dc4afd69c110c
work_keys_str_mv	AT oktaniasandrapuspita optimizingrealtimepcrmethodsfordetectionofssangenesalmonellaentericasubspentericainthebloodspecimen AT andiyasmon optimizingrealtimepcrmethodsfordetectionofssangenesalmonellaentericasubspentericainthebloodspecimen AT betiernawatidewi optimizingrealtimepcrmethodsfordetectionofssangenesalmonellaentericasubspentericainthebloodspecimen
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Optimizing Real-Time PCR methods for detection of ssaN gene Salmonella enterica subsp.enterica in the blood specimen

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