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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.
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Background: Malawi is among 7 countries participating in the Enterics for Global Health (EFGH) Shigella surveillance study, which aims to determine the incidence of medically attended diarrhea attributed to Shigella, a leading bacterial cause of diarrhea in children in low-resource settings. Methods: We describe the EFGH study site in the densely populated informal settlement of Ndirande Township, Blantyre, Malawi. We explore the site's geographical location, demographic characteristics, and the healthcare-seeking behavior of its population, particularly for childhood diarrhea. We also describe the management of childhood diarrhea at the health facility, and the associated challenges to attaining optimum adherence to local and national guidelines at the site. Conclusions: Our overarching aim is to improve global health through understanding and mitigating the impact of diarrhea attributed to Shigella.
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Introduction: Despite global evidence of chikungunya fever (CHIKF) in humans that is caused by chikungunya virus (CHIKV), little is known about the occurrence of CHIKF in Malawi. This study was conducted to determine the seroprevalence of CHIKF and to molecularly confirm the presence of CHIKV ribonucleic acid (RNA) among febrile outpatients seeking health care at Mzuzu Central Hospital in the Northern Region of Malawi. Methods: Enzyme-immunosorbent assay (ELISA) was used to detect the presence or absence of specific antibodies against CHIKV. Reversetranscription polymerase chain reaction (RT-PCR) was conducted on randomly selected anti-CHIKV IgM-positive samples to detect CHIKV RNA. Results: Out of 119 CHIKF suspected samples analyzed, 73 tested positive for anti-CHIKV IgM antibodies, with an overall seroprevalence of 61.3%. Most of the CHIKV infected individuals presented with joint pain, abdominal pain, vomiting and nose bleeding with seroprevalence of 45.2%, 41.1%, 16.4% and 12.3%, respectively. All the randomly selected samples that were positive for CHIKV anti-IgM by ELISAhad detectable CHIKV RNA by RT-PCR. Conclusion: The presence of anti-CHIKV IgM antibodies suggests the presence of recent CHIKV infection. We therefore recommend for the inclusion of CHIKF as the differential diagnosis in febrile ill patients in Mzuzu city, Malawi.
