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Table 1.

Primer and probe sequences.

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Table 2.

Salmonella Typhi antigens for ELISA confirmation of blood culture negative/ PCR positive samples.

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Fig 1.

Flowchart of participant recruitment.

Numbers of children screened, recruited and processed on PCR, blood culture and serology are provided.

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Table 3.

Characteristics of 643 children ≤ 4 years.

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Fig 2.

Bimodal distribution of CT-values for primers invA, fliC and staG.

Graph shows the distribution of CT-values for the salmonella pan primer and S.Typhi and S.Typhiurium specific primers with a clear distinction between PCR positives and negatives.

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Fig 3.

CT cut-off set to demarcate non-specific amplification signals for primers invA (a), fliC (b) and staG (c). Graph shows CT-values from serial dilutions of contaminants; Micrococci (mc), Bacillus (bac), and pathogens; S. Typhi (sty), S. Typhimurium (stm), S. aureus (sta), Klebsiella (klb), and E. coli (eco).

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Table 4.

Blood culture growth observed in the study population.

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Table 5.

PCR performance in relation to Blood culture.

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Fig 4.

Distribution of IgG antibody titres during acute infection (A, C, E, G) and in convalescence (B, D, F, H). IgG antibody titres were measured against four antigens (STY1498 in A & B, STY1479 in C & D, STY1886 in E & F, & Vi in G & H) across distinct groups of children. The groups are bc+PCR+ (cases that had S. Typhi infection confirmed by both blood culture and PCR (or blood culture alone), in acute infection [age 14 to 45, median 37 months], and in convalescence [age 14 to 45, median 33 months]), PCR+ (S. Typhi infection confirmed by PCR only, in cute infection [age 8 to 46, median 22 months] and in convalescence [age 22 to 46, median 40 months]), Negative (febrile but negative for typhoid on blood culture and PCR, in acute infection [age 1 to 48, median 13.5 months] and in convalescence [age 1 to 45, median 16 months]), and Control (afebrile healthy controls [age 0 to 52.5, median 10.2 months]).

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Fig 5.

Distribution of IgM antibody titres during acute infection (A, C, E, G) and in convalescence (B, D, F, H). IgM antibodies were measured against four antigens (STY1498 in A & B, STY1479 in C & D, STY1886 in E & F & Vi in G &H) across distinct groups of children. The groups are bc+PCR+ (cases that had S. Typhi infection confirmed by both blood culture and PCR (or blood culture alone), in acute infection [age 14 to 45, median 37 months], and in convalescence [age 14 to 45, median 33 months]), PCR+ (S. Typhi infection confirmed by PCR only, in cute infection [age 8 to 46, median 22 months] and in convalescence [age 22 to 46, median 40 months]), Negative (febrile but negative for typhoid on blood culture and PCR, in acute infection [age 1 to 48, median 13.5 months] and in convalescence [age 1 to 45, median 16 months]), and Control (afebrile healthy controls [age 0 to 52.5, median 10.2 months]).

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Fig 5 Expand