Simultaneous detection of chikungunya virus, dengue virus and human pathogenic Leptospira genomes using a multiplex TaqMan® assay

BMC Microbiology

Methodology Article

Claude Giry¹  Ghislaine Li-Pat-Yuen¹  Marie-Christine Jaffar-Bandjee²  Bénédicte Roquebert³  Philippe Gasque⁴ 

[1] Centre National Arbovirus Associé, CHU de la Réunion-Site Nord, Saint-Denis, Réunion, France;Laboratoire de microbiologie et, CHU de la Réunion-Site Nord, Saint-Denis, Réunion, France;Centre National Arbovirus Associé, CHU de la Réunion-Site Nord, Saint-Denis, Réunion, France;Laboratoire de microbiologie et, CHU de la Réunion-Site Nord, Saint-Denis, Réunion, France;Laboratoire d’immunologie clinique et expérimentale ZOI (LICE-OI), CHU de la Réunion-Site Nord, Saint-Denis, Réunion, France;UMR PIMIT, Processus Infectieux en Milieu Insulaire Tropical, Université de la Réunion, INSERM 1187, CNRS 9192, IRD 249, Saint-Denis, Réunion, France;Centre National Arbovirus Associé, CHU de la Réunion-Site Nord, Saint-Denis, Réunion, France;Laboratoire de microbiologie et, CHU de la Réunion-Site Nord, Saint-Denis, Réunion, France;UMR PIMIT, Processus Infectieux en Milieu Insulaire Tropical, Université de la Réunion, INSERM 1187, CNRS 9192, IRD 249, Saint-Denis, Réunion, France;Laboratoire d’immunologie clinique et expérimentale ZOI (LICE-OI), CHU de la Réunion-Site Nord, Saint-Denis, Réunion, France;UMR PIMIT, Processus Infectieux en Milieu Insulaire Tropical, Université de la Réunion, INSERM 1187, CNRS 9192, IRD 249, Saint-Denis, Réunion, France;

关键词: Chikungunya;    Dengue;    Leptospira;    Multiplex real-time RT-PCR;   

DOI  :  10.1186/s12866-017-1019-1

 received in 2017-01-26, accepted in 2017-04-28,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundIn 2005–2006 a major epidemics of Chikungunya disease occurred in South-West Indian Ocean islands. In Reunion Island, the magnitude of Chikungunya infection related symptoms was high and with over 38% of serological prevalence in the population. This epidemics illustrated the potential threat of emerging arboviral diseases for inhabitants of Reunion Island and elsewhere since vectors are worldwide distributed. A sentinel surveillance network was set-up to detect emerging pathogens associated with fever over 38 °C and in the absence of known etiologic causes. Leptospirosis is caused by a pathogenic spirochete of the Leptospira genus and is an endemic and recurrent seasonal disease of great concern in Reunion Island. To accurately diagnose potentially infected patients and to advise Health authorities on the presence of emerging pathogens, a rapid diagnostic test was needed that could differentiate between these 3 pathogens.MethodsA one-step multiplex real-time PCR assay was developed that can simultaneously detect RNA of Chikungunya and Dengue viruses and leptospiral DNA with good performance for a routine diagnostic use.ResultsSimplex protocols already published were used with key modifications to implement a triplex assay which was set-up with a small reaction volume to improve cost efficiency.ConclusionsThis approach has enabled greater diagnostic capacity in our laboratory. We established a multiplex approach validated and valuable for cost savings, and with the concurrent detection of 3 pathogens of public health concern.

【 授权许可】

CC BY   
© The Author(s). 2017