Simultaneous and high sensitive detection of Salmonella typhi and Salmonella paratyphi a in human clinical blood samples
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Simultaneous and high sensitive detection of Salmonella typhi and Salmonella paratyphi a in human clinical blood samples using an affordable and portable device Avinash Kaur 1 & Ankur Ruhela 1 & Priyanka Sharma 2 & Harshit Khariwal 3 & Sagar Seth 4 & Adarsh Kumar 3 & Arti Kapil 2 & Ravikrishnan Elangovan 5 & Dinesh Kalyanasundaram 1,6
# Springer Science+Business Media, LLC, part of Springer Nature 2019
Abstract Enteric fever is one of the leading causes of infection and subsequent fatality (greater than 1.8 million) (WHO 2018), especially in the developing countries due to contaminated water and food inter twinned with unhygienic practices. Clinical gold standard technique of culture-based method followed by biochemical tests demand 72+ hours for diagnosis while newly developed techniques (like PCR, RT-PCR, DNA microarray etc.) suffer from high limit of detection or involve high-cost infrastructure or both. In this work, a quick and highly specific method, SMOL was established for simultaneous detection of Salmonella paratyphi A and Salmonella typhi in clinical blood samples. SMOL consists of (i) pre-concentration of S. typhi and S. paratyphi A cells using magnetic nanoparticles followed by (ii) cell lysis and DNA extraction (iii) amplification of select nucleic acids by LAMP technique and (iv) detection of amplified nucleic acids using an affordable portable device (costs less than $70). To identify the viability of target cells at lower concentrations, the samples were processed at two different time periods of t = 0 and t = 4 h. Primers specific for the SPA2539 gene in S. paratyphi A and STY2879 gene in S. typhi were used for LAMP. Within 6 h SMOL was able to detect positive and negative samples from 55 human clinical blood culture samples and detect the viability of the cells. The results were concordant with culture and biochemical tests as well as by qPCR. Statistical power analysis yielded 100%. SMOL results were concordant with culture and biochemical tests as well as by qPCR. The sensitive and affordable system SMOL will be effective for poor resource settings. Keywords Salmonella typhi . Salmonella paratyphi A . LAMP . Diagnosis . Cross-reactivity . Device
1 Introduction Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10544-019-0441-6) contains supplementary material, which is available to authorized users. * Dinesh Kalyanasundaram [email protected]; [email protected] 1
Centre for Biomedical Engineering, Indian Institute of Technology Delhi, New Delhi 110016, India
2
Department of Microbiology, All India Institute of Medical Sciences, New Delhi 110029, India
3
Department of Electrical Engineering, Indian Institute of Technology Bombay, Mumbai 400076, India
4
Department of Physics, Indian Institute of Technology Bombay, Mumbai 400076, India
5
Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, New Delhi 110016, India
6
Department of Biomedical Engineering, All India Institute of Medical Sc
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