Shigella Detection and Molecular Serotyping With a Customized TaqMan Array Card in the Enterics for Global Health (EFGH): Shigella Surveillance Study

• Published in: Open forum infectious diseases Vol. 11; no. Supplement_1; pp. S34 - S40
• Main Authors: Liu, Jie, Garcia Bardales, Paul F, Islam, Kamrul, Jarju, Sheikh, Juma, Jane, Mhango, Chimwemwe, Naumanga, Queen, Qureshi, Sonia, Sonye, Catherine, Ahmed, Naveed, Aziz, Fatima, Bhuiyan, Md Taufiqur Rahman, Charles, Mary, Cunliffe, Nigel A, Abdou, Mahamadou, Galagan, Sean R, Gitteh, Ensa, Guindo, Ibrehima, Jahangir Hossain, M, Jabang, Abdoulie M J, Jere, Khuzwayo C, Kawonga, Flywell, Keita, Mariama, Keita, Noumou Yakhouba, Kotloff, Karen L, Shapiama Lopez, Wagner V, Munga, Stephen, Paredes Olortegui, Maribel, Omore, Richard, Pavlinac, Patricia B, Qadri, Firdausi, Qamar, Farah Naz, Azadul Alam Raz, S M, Riziki, Laura, Schiaffino, Francesca, Stroup, Suzanne, Traore, Sarata Nassoun, Pinedo Vasquez, Tackeshy, Yousafzai, Mohammad Tahir, Antonio, Martin, Cornick, Jennifer E, Kabir, Furqan, Khanam, Farhana, Kosek, Margaret N, Ochieng, John Benjamin, Platts-Mills, James A, Tennant, Sharon M, Houpt, Eric R
• Format: Journal Article
• Language: English
• Published: US Oxford University Press 25-03-2024
• Subjects: Supplement; serotyping; quantification; PCR; rectal swab; ipaH
• ISSN: 2328-8957; 2328-8957
• DOI: 10.1093/ofid/ofad574

Abstract
Background
Quantitative polymerase chain reaction (qPCR) targeting ipaH has been proven to be highly efficient in detecting Shigella in clinical samples compared to culture-based methods, which underestimate Shigella burden by 2- to 3-fold. qPCR assays have also been developed for Shigella speciation and serotyping, which is critical for both vaccine development and evaluation.
Methods
The Enterics for Global Health (EFGH) Shigella surveillance study will utilize a customized real-time PCR–based TaqMan Array Card (TAC) interrogating 82 targets, for the detection and differentiation of Shigella spp, Shigella sonnei, Shigella flexneri serotypes, other diarrhea-associated enteropathogens, and antimicrobial resistance (AMR) genes. Total nucleic acid will be extracted from rectal swabs or stool samples, and assayed on TAC. Quantitative analysis will be performed to determine the likely attribution of Shigella and other particular etiologies of diarrhea using the quantification cycle cutoffs derived from previous studies. The qPCR results will be compared to conventional culture, serotyping, and phenotypic susceptibility approaches in EFGH.
Conclusions
TAC enables simultaneous detection of diarrheal etiologies, the principal pathogen subtypes, and AMR genes. The high sensitivity of the assay enables more accurate estimation of Shigella-attributed disease burden, which is critical to informing policy and in the design of future clinical trials.
Shigella spp, Shigella flexneri serotypes, and other diarrhea-associated enteropathogens are detected and quantified with a customized real-time polymerase chain reaction–based TaqMan Array Card in the Enterics for Global Health: Shigella surveillance study. Shigella burden will be estimated with a quantitative methodology.