Systematic application of multiplex PCR enhances the detection of bacteria, parasites, and viruses in stool samples

McAuliffe, Gary; Anderson, Trevor P.; Stevens, Mary Betty; Adams, J. W.; Coleman, Robyn; Mahagamasekera, Patalee; Young, Sheryl A.; Henderson, Tom; Hofmann, Maria; Jennings, Lance C.; Murdoch, David R.

doi:10.1016/j.jinf.2013.04.009

Cited by 93 publications

(75 citation statements)

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“…Not only are these assays more sensitive than routine culture and EIA methods, they are also more likely to detect coinfections, as evidenced in this evaluation and others (6,10,21). However, multiplex assays that report bacterial, viral, and parasitic targets simultaneously may generate results that were not requested by clinicians and create complexities in reporting as well as in interpretation (17,29,30). For example, detection of Clostridium difficile in colonized individuals who actually harbor an agent of acute bacterial enteritis may lead to overtreatment for this pathogen.…”

Section: Discussionmentioning

confidence: 78%

“…The BD Max EBP assay is performed with the BD Max instrument, a walkaway instrument that conducts all steps of the molecular assay from extraction to interpretation of results. Compared to routine stool culture, laboratories have demonstrated significant reductions in turnaround time with the use of molecular assays (17,18). Additionally, the CDC recommends culture-based screening for E. coli O157 and EIA or molecular methods for Shiga toxins (19).…”

Section: Discussionmentioning

confidence: 99%

“…in the prospective arm, there was little difference in performance between prospective and retrospective data sets (Table 3). Reduced sensitivity for Salmonella in comparison to culture has been reported (12,17,21,25). Cunningham et al attributed reduced sensitivity to the enhancement of the culture method with a selenite enrichment broth (12).…”

Section: Discussionmentioning

confidence: 99%

“…The ability to test either specimen type may allow better flexibility in specimen collection practices than those required for some other methods. One potential weakness is the comparison to culture and EIA methods, which are known to lack sensitivity for enteric pathogens (6,9,12,17). However, the use of alternate PCR and bidirectional sequencing allows a more direct comparison of similar methods for specimens that gave discrepant results.…”

Section: Discussionmentioning

confidence: 99%

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Multicenter Evaluation of the BD Max Enteric Bacterial Panel PCR Assay for Rapid Detection of Salmonella spp., Shigella spp., Campylobacter spp. (C. jejuni and C. coli), and Shiga Toxin 1 and 2 Genes

et al. 2015

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i Diarrhea due to enteric bacterial pathogens causes significant morbidity and mortality in the United States and worldwide. However, bacterial pathogens may be infrequently identified. Currently, culture and enzyme immunoassays (EIAs) are the primary methods used by clinical laboratories to detect enteric bacterial pathogens. We conducted a multicenter evaluation of the BD Max enteric bacterial panel (EBP) PCR assay in comparison to culture for the detection of Salmonella spp., Shigella spp., Campylobacter jejuni, and Campylobacter coli and an EIA for Shiga toxins 1 and 2. A total of 4,242 preserved or unpreserved stool specimens, including 3,457 specimens collected prospectively and 785 frozen, retrospective samples, were evaluated. Compared to culture or EIA, the positive percent agreement (PPA) and negative percent agreement (NPA) values for the BD Max EBP assay for all specimens combined were as follows: 97.1% and 99.2% for Salmonella spp., 99.1% and 99.7% for Shigella spp., 97.2% and 98.4% for C. jejuni and C. coli, and 97.4% and 99.3% for Shiga toxins, respectively. Discrepant results for prospective samples were resolved with alternate PCR assays and bidirectional sequencing of amplicons. Following discrepant analysis, PPA and NPA values were as follows: 97.3% and 99.8% for Salmonella spp., 99.2% and 100% for Shigella spp., 97.5% and 99.0% for C. jejuni and C. coli, and 100% and 99.7% for Shiga toxins, respectively. No differences in detection were observed for samples preserved in Cary-Blair medium and unpreserved samples. In this large, multicenter study, the BD Max EBP assay showed superior sensitivity compared to conventional methods and excellent specificity for the detection of enteric bacterial pathogens in stool specimens. The World Health Organization reports that, worldwide, there are nearly 1.7 billion cases of diarrheal disease every year and that diarrheal disease is the second leading cause of death of children Ͻ5 years old. Each year, diarrhea kills ϳ760,000 children under the age of 5 years, and most importantly, it is preventable and treatable. Diarrhea is also a leading cause of malnutrition in this same age group. Most of this disease is related to unsafe drinking water, inadequate sanitation, and poor hygiene (1, 2).Acute gastroenteritis is caused by a number of bacterial, viral, and parasitic agents (3). In the United States, noroviruses cause most cases of self-limited, acute gastroenteritis. However, Salmonella, Campylobacter, Shigella, and Shiga toxin-producing Escherichia coli (STEC) are the most common diarrheagenic bacteria, and they are usually associated with foodborne illness (4). Furthermore, Shigella is more frequently transmitted from person to person due to the low infectious dose. Importantly, agents of gastroenteritis may not be distinguished clinically. Identifying the cause of diarrhea is important for both the treatment of individual patients and public health intervention through outbreak management (5).Conventional microbiological cultures remain the gold standard...

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Section: Discussionmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Multicenter Evaluation of the BD Max Enteric Bacterial Panel PCR Assay for Rapid Detection of Salmonella spp., Shigella spp., Campylobacter spp. (C. jejuni and C. coli), and Shiga Toxin 1 and 2 Genes

et al. 2015

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“…Сравнительные исследования, проведенные разными авторами, так же как и наши данные, доказывают, что ПЦР значительно эффектив-нее для выявления любого возбудителя диареи, чем традиционные бактериологические методы [2,5,9,12,14]. Особенно диагностическое пре-восходство ПЦР значимо в выявлении таких возбудителей ОКИ, как Campylobacter spp., E. coli и Clostridium difficile.…”

Section: Discussionunclassified

Acute Enteric Infections Polymerase Chain Reaction Assay in Pediatric Practice: Opportunities and Challenges

Дмитриевна

Галтаева

Замурий

et al. 2016

Russian Journal of Infection and Immunity

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Резюме. Изучена этиологическая структура острых кишечных инфекций у детей, госпитализированных с диа-реей в детский инфекционный стационар, с целью оценки возможностей совершенствования этиологической расшифровки заболеваний при дополнении традиционных бактериологических методов ПЦР-диагностикой с применением отечественных мульти-прайм ПЦР-реагентов. Использовали 4 набора реагентов, обеспечиваю-щих одновременную индикацию нескольких патогенов в одной пробе: 1) Rotavirus, Norovirus, Astrovirus; 2) Shigella spp./EIEC, Salmonella spp., Campylobacter spp.; 3) Yersinia enterocolitica и Yersinia pseudotuberculosis; 4) Escherichia coli: EIEC (энтероинвазивные), EPEC (энтеропатогенные), ETEC (энтеротоксигенные), EHEC (энтерогеморраги-ческие), EAgEC (энтероаггрегативные). Показано, что этиологическая диагностика вирусных диарей увели-чивается на 14%, бактериальных -в 2,5 раза. ПЦР-диагностика выявила у 62% пациентов вирусную природу гастроэнтеритов: Rotavirus (52%), Norovirus (9%), Astrovirus (1%). Количество выявленных ПЦР-маркеров бак-териальных возбудителей ОКИ оказалось в 2,5 раза больше, чем по данным бактериологического исследова-ния. Спектр выявляемых бактериальных агентов увеличился за счет E. coli и Y. enterocolitica. ПЦР-диагностика в 2 раза повысила выявляемость Campylobacter spp. и оказалась единственно эффективным методом детекции ДНК E. сoli в пробах испражнений: доминировали EPEC -66%, EAgEC, ETEC и EHEC составили 31, 9 и 4% соответственно, которые не могли быть идентифицированы при бактериологическом исследовании. ДНК Campylobacter spp. и E. coli в сумме составляли 2 / 3 находок среди бактериальных возбудителей: Campylobacter spp. (41%), E. coli (24%), Salmonella spp. (19%), Yersinia spp. (11%), Shigella spp./EIEC (5%). Положительные результаты бактериологического посева проб испражнений и серологического исследования крови в РНГА дублировали положительные результаты ПЦР-диагностики. В целом, положительные результаты ПЦР-диагностики бак-териальных патогенов установлены у 46,35% обследованных пациентов. У 53,65% пациентов выявлены ПЦР-маркеры ОКИ смешанной этиологии, в основном вирусно-бактериальной, из них в 48,4% случаях выявлены 19 различных вариантов ассоциаций. 12 вариантов бактериальных ассоциаций обнаружены в 5,25%, из них у 11% обнаружены ДНК 2-3 возбудителей бактериальных ОКИ. Проведенное исследование расширило наши Адрес для переписки: Соколова Екатерина Дмитриевна 192289, Санкт-Петербург, ул. Бухарестская, 134, Детская городская клиническая больница № 5 им. Н.Ф. Филатова.

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Parasitic Diseases: Protozoa

Stanley

2015

Yamada' S Textbook of Gastroenterology

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Systematic application of multiplex PCR enhances the detection of bacteria, parasites, and viruses in stool samples

Cited by 93 publications

References 30 publications

Multicenter Evaluation of the BD Max Enteric Bacterial Panel PCR Assay for Rapid Detection of Salmonella spp., Shigella spp., Campylobacter spp. (C. jejuni and C. coli), and Shiga Toxin 1 and 2 Genes

Multicenter Evaluation of the BD Max Enteric Bacterial Panel PCR Assay for Rapid Detection of Salmonella spp., Shigella spp., Campylobacter spp. (C. jejuni and C. coli), and Shiga Toxin 1 and 2 Genes

Acute Enteric Infections Polymerase Chain Reaction Assay in Pediatric Practice: Opportunities and Challenges

Parasitic Diseases: Protozoa

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