Travelers’ Diarrhea in Thailand: A Quantitative Analysis Using TaqMan® Array Card

Lertsethtakarn, Paphavee; Silapong, Sasikorn; Sakpaisal, Pimmada; Serichantalergs, Oralak; Ruamsap, Nattaya; Lurchachaiwong, Woradee; Anuras, Sinn; Platts-Mills, James A; Liu, Jie; Houpt, Eric R.; Bodhidatta, Ladaporn; Swierczewski, Brett E.; Mason, Carl J.

doi:10.1093/cid/ciy040

Cited by 36 publications

(56 citation statements)

References 39 publications

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“…In a recent report, travelers' diarrhea in foreign visitors to Thailand was investigated using a TaqMan array card assay 30 . One hundred and seventy-three cases from the in-patient or out-patient department in a private hospital and 165 non-diarrheal subjects were enrolled in that study.…”

Section: Discussionmentioning

confidence: 99%

Etiologic features of diarrheagenic microbes in stool specimens from patients with acute diarrhea in Thailand

Okada

Wongboot

Kamjumphol³

et al. 2020

Sci Rep

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Many microbial species have been recognized as enteropathogens for humans. Here, we predicted the causative agents of acute diarrhea using data from multiplex quantitative PCR (qPCR) assays targeting 19 enteropathogens. For this, a case-control study was conducted at eight hospitals in Thailand. Stool samples and clinical data were collected from 370 hospitalized patients with acute diarrhea and 370 non-diarrheal controls. Multiple enteropathogens were detected in 75.7% and 13.0% of diarrheal stool samples using multiplex qPCR and bacterial culture methods, respectively. Asymptomatic carriers of enteropathogens were found among 87.8% and 45.7% of individuals by qPCR and culture methods, respectively. These results suggested the complexity of identifying causative agents of diarrhea. An analysis using the quantification cutoff values for clinical relevance drastically reduced pathogenpositive stool samples in control subjects from 87.8% to 0.5%, whereas 48.9% of the diarrheal stool samples were positive for any of the 11 pathogens. Among others, rotavirus, norovirus GII, Shigella/ EIEC, and Campylobacter were strongly associated with acute diarrhea (P-value < 0.001). Characteristic clinical symptoms, epidemic periods, and age-related susceptibility to infection were observed for some enteropathogens. Investigations based on qPCR approaches covering a broad array of enteropathogens might thus improve our understanding of diarrheal disease etiology and epidemiological trends. Diarrheal diseases are one of the major causes of mortality and morbidity worldwide, especially during the first 5 years of life for individuals subjected to malnutrition 1-3. Diarrhea can be defined by increased stool frequency, liquidity, or volume 4. A wide range of enteropathogens including bacteria, viruses, and protozoa have been

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Section: Discussionmentioning

confidence: 99%

Etiologic features of diarrheagenic microbes in stool specimens from patients with acute diarrhea in Thailand

Okada

Wongboot

Kamjumphol³

et al. 2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…We conducted a pilot study using a customized TD TaqMan Array Card (TAC) to assess the feasibility of using archived diarrheal specimens, of varying time periods, focusing on four TD pathogens: enterotoxigenic E coli (ETEC), Shigella spp., Campylobacter spp., and Norovirus. Two Department of Defense (DoD) fecal repositories of adult TD cases, tested using standard microbiologic methods at the time of sample collection, were utilized for the study [7,8] We evaluated the Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) of TAC for detecting TD pathogens in archived fecal specimens, using results from previous microbiologic testing as the 'benchmark', the impact of storage duration and use of FTA cards on TAC sensitivity. Approximately 65 positive stool specimens per pathogen for ETEC, Shigella spp., Campylobacter spp., and Norovirus (based on an estimated PPA and NPA of 80% for the TAC assay and precision of 10%) and 30 samples negative for all pathogens, were requested.…”

Section: Methodsmentioning

confidence: 99%

Performance Characteristics of the TaqMan Array PCR on Repository Stool Specimens and Smears on Whatman® FTA Elute Cards

Tisdale¹,

Mitra²,

McCoy³

et al. 2020

Preprint

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Objective Stool repositories are a valuable resource for retrospective analyses including quantitative PCR assays to distinguish between asymptomatic shedding and clinical disease. The suitability of archival specimens for this purpose is unclear and requires assessment. We conducted a pilot study to evaluate pathogen detection by TaqMan Array Card (TAC) in travelers’ diarrhea (TD) stool specimens stored for 1–13 years, as well as, the impact of transporting specimens on Whatman FTA Elute cards (FTA Cards) on detection. Results The positive percent agreement (PPA) for TAC on stool vs. microbiologic testing was lower than our a priori PPA estimate of 80% for most pathogens: Shigella spp. (100% [95%CI: 69–100%]), enterotoxigenic E coli (ETEC) (63 [49–75%]), Campylobacter spp. (66 [43–85%]) and Norovirus (37 [16–61%]). Use of the FTA card resulted in a further reduction of PPA. Our findings suggest that archival specimens may lead to insensitive detection on quantitative PCR assays due to degradation of nucleic acid with prolonged storage, although our limited sample size precluded us from evaluating the impact of storage duration on nucleic acid yield. Additional studies are needed to understand the impact of storage duration on quantitative PCR data.

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“…We conducted a pilot study using a customized TD TaqMan Array Card (TAC, Life Technologies, Carlsbad, CA, USA) to assess the feasibility of using archived diarrheal specimens, of varying time periods, focusing on four TD pathogens: enterotoxigenic E coli (ETEC), Shigella spp., Campylobacter spp., and Norovirus. Two United States Department of Defense (DoD) fecal repositories of adult TD cases, tested using standard microbiologic methods at the time of sample collection, were utilized for the study [7,8]:…”

Section: Methodsmentioning

confidence: 99%

Performance characteristics of a quantitative PCR assay on repository stool specimens and smeared filter-paper cards

Tisdale¹,

Mitra

McCoy

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Objective Stool repositories are a valuable resource for retrospective analyses including quantitative PCR assays to distinguish between asymptomatic shedding and clinical disease. The suitability of archival specimens for this purpose is unclear and requires assessment. We conducted a pilot study to evaluate pathogen detection by TaqMan Array Card (TAC) in travelers’ diarrhea (TD) stool specimens stored for 1-13 years, as well as the impact of transporting specimens on Whatman FTA Elute cards (FTA Cards) on detection. Results The positive percent agreement (PPA) for TAC on stool vs. microbiologic testing was lower than our a priori PPA estimate of 80% for most pathogens: Shigella spp. (100% [95%CI: 69-100%]), enterotoxigenic E coli (ETEC) (63% [95%CI: 49-75%]), Campylobacter spp. (66% [95%CI: 43-85%]) and Norovirus (37% [95%CI: 16-61%]). Use of the FTA card resulted in a further reduction of PPA. Our findings suggest that archival specimens may lead to insensitive detection on quantitative PCR assays due to degradation of nucleic acid with prolonged storage, although our limited sample size precluded us from evaluating the impact of storage duration on nucleic acid yield. Additional studies are needed to understand the impact of storage duration on quantitative PCR data.

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Travelers’ Diarrhea in Thailand: A Quantitative Analysis Using TaqMan® Array Card

Cited by 36 publications

References 39 publications

Etiologic features of diarrheagenic microbes in stool specimens from patients with acute diarrhea in Thailand

Etiologic features of diarrheagenic microbes in stool specimens from patients with acute diarrhea in Thailand

Performance Characteristics of the TaqMan Array PCR on Repository Stool Specimens and Smears on Whatman® FTA Elute Cards

Performance characteristics of a quantitative PCR assay on repository stool specimens and smeared filter-paper cards

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