TY - JOUR
T1 - Evaluation of a simple, rapid and field-adapted diagnostic assay for enterotoxigenic E. coli and Shigella
AU - Connor, Sean
AU - Velagic, Mirza
AU - Zhang, Xueyan
AU - Johura, Fatema Tuz
AU - Chowdhury, Goutam
AU - Mukhopadhyay, Asish K.
AU - Dutta, Shanta
AU - Alam, Munirul
AU - Sack, David A.
AU - Wierzba, Thomas F.
AU - Chakraborty, Subhra
N1 - Publisher Copyright:
© 2022 Connor et al.
PY - 2022/2
Y1 - 2022/2
N2 - Understanding the global burden of enterotoxigenic E. coli (ETEC) and Shigella diarrhea as well as estimating the cost effectiveness of vaccines to control these two significant pathogens have been hindered by the lack of a diagnostic test that is rapid, simple, sensitive, and can be applied to the endemic countries. We previously developed a simple and rapid assay, Rapid Loop mediated isothermal amplification based Diagnostic Test (RLDT) for the detection of ETEC and Shigella spp. (Shigella). In this study, the RLDT assay was evaluated in comparison with quantitative PCR (qPCR), culture and conventional PCR for the detection of ETEC and Shigella. This validation was performed using previously collected stool samples from endemic countries, from the travelers to the endemic countries, as well as samples from a controlled human infection model study of ETEC. The performance of RLDT from dried stool spots was also validated. RLDT resulted in excellent sensitivity and specificity compared to qPCR (99% and 99.2% respectively) ranging from 92.3 to 100% for the indi-vidual toxin genes of ETEC and 100% for Shigella. Culture was less sensitive compared to RLDT. No significant differences were noted in the performance of RLDT using samples from various sources or stool samples from moderate to severe diarrhea or asymptomatic infections. RLDT performed equally well in detection of ETEC and Shigella from the dried stool samples on filter papers. This study established that RLDT is sufficiently sensitive and specific to be used as a simple and rapid diagnostic assay to detect ETEC and Shigella in endemic countries to determine disease burden of these pathogens in the national and sub-national levels. This information will be important to guide public health and policy makers to prioritize resources for accelerating the development and introduction of effective preventa-tive and/or treatment interventions against these enteric infections.
AB - Understanding the global burden of enterotoxigenic E. coli (ETEC) and Shigella diarrhea as well as estimating the cost effectiveness of vaccines to control these two significant pathogens have been hindered by the lack of a diagnostic test that is rapid, simple, sensitive, and can be applied to the endemic countries. We previously developed a simple and rapid assay, Rapid Loop mediated isothermal amplification based Diagnostic Test (RLDT) for the detection of ETEC and Shigella spp. (Shigella). In this study, the RLDT assay was evaluated in comparison with quantitative PCR (qPCR), culture and conventional PCR for the detection of ETEC and Shigella. This validation was performed using previously collected stool samples from endemic countries, from the travelers to the endemic countries, as well as samples from a controlled human infection model study of ETEC. The performance of RLDT from dried stool spots was also validated. RLDT resulted in excellent sensitivity and specificity compared to qPCR (99% and 99.2% respectively) ranging from 92.3 to 100% for the indi-vidual toxin genes of ETEC and 100% for Shigella. Culture was less sensitive compared to RLDT. No significant differences were noted in the performance of RLDT using samples from various sources or stool samples from moderate to severe diarrhea or asymptomatic infections. RLDT performed equally well in detection of ETEC and Shigella from the dried stool samples on filter papers. This study established that RLDT is sufficiently sensitive and specific to be used as a simple and rapid diagnostic assay to detect ETEC and Shigella in endemic countries to determine disease burden of these pathogens in the national and sub-national levels. This information will be important to guide public health and policy makers to prioritize resources for accelerating the development and introduction of effective preventa-tive and/or treatment interventions against these enteric infections.
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U2 - 10.1371/journal.pntd.0010192
DO - 10.1371/journal.pntd.0010192
M3 - Article
C2 - 35130310
AN - SCOPUS:85124850669
SN - 1935-2727
VL - 16
JO - PLoS neglected tropical diseases
JF - PLoS neglected tropical diseases
IS - 2
M1 - e0010192
ER -