Evaluation of a multiplex PCR assay for simultaneous detection of bacterial and viral enteropathogens in stool samples of paediatric patients

Onori, Manuela; Coltella, Luana; Mancinelli, Livia; Argentieri, Marta; Menichella, D.; Villani, Alberto; Grandin, Annalisa; Valentini, Diletta; Raponi, Massimiliano; Russo, Cristina

doi:10.1016/j.diagmicrobio.2014.02.004

Cited by 73 publications

(49 citation statements)

References 28 publications

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“…We found that, when co-infections were tested for in children with a positive C. difficile test, a variety of pathogens were encountered (Table 2). This pattern is not specific to children with a positive C. difficile test and can be expected in any child with diarrhoea [44]. Unfortunately, only four studies systematically tested for the presence of viruses, bacteria and parasites, and the specific pathogen was not reported in over one-third of co-infections, which limits the reliability of the data and underlines the need for future studies.…”

Section: Discussionmentioning

confidence: 59%

“…In those that did, the panel of co-infections tested for varied considerably. Taking this into account, along with the fact that a causative organism is found in only 23.2-67 % of children with diarrhoea [44,45], this review highlights an under-appreciation of coinfections in children, and the true rate could be substantially higher than the reported pooled rate. Unfortunately, the studies in our analysis often consisted of small cohorts (with a mean of 55 C. difficile-positive samples), were not stratified by age groups or risk factors, and outcomes such as survival, length of hospital stay and incidence of complications were not discussed in depth, preventing a more meaningful interpretation of the data.…”

Section: Discussionmentioning

confidence: 91%

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Co-infection as a confounder for the role of Clostridium difficile infection in children with diarrhoea: a summary of the literature

Pai

Burns

Karas

et al. 2015

Eur J Clin Microbiol Infect Dis

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Although Clostridium difficile is a major cause of antibiotic-associated diarrhoea in adults, the incidence and severity of C. difficile infection (CDI) in children is unclear. One complicating factor in assessing the role of CDI in children is the possibility of co-infection with other gastrointestinal pathogens. In this review, we summarise the literature concerning C. difficile co-infections in young children, in an attempt to discuss the rate of co-infections and their potential role in the severity of CDI clinical presentation. We identified 31 studies where co-infections were analysed, comprising 1,718 patients with positive C. difficile tests. The pooled percentage of reported co-infections was 20.7% (range 0-100%). Viral co-infections were most commonly reported (46%), with bacteria and parasites accounting for 14.9% and 0.01% of cases, respectively. However, the panel of co-infections tested for varied considerably among studies and 38% of stated co-infections did not have a pathogen reported. Substantial variation in how and when tests for gastrointestinal co-infections are carried out, small sample sizes and a lack of clear CDI case definitions preclude meaningful conclusions on the true rate of co-infections in this patient population. This review suggests that co-infections may be common in children with diarrhoea who tested positive for C. difficile. Given a lack of CDI case definitions, especially in young children under the age of 5 years, a broad panel of pathogens should be tested for to exclude other microbiological causes. However, the summarised poor quality of the available literature on this subject highlights a need for further studies.

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Section: Discussionmentioning

confidence: 59%

Section: Discussionmentioning

confidence: 91%

Co-infection as a confounder for the role of Clostridium difficile infection in children with diarrhoea: a summary of the literature

Pai

Burns

Karas

et al. 2015

Eur J Clin Microbiol Infect Dis

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“…of the positive samples. The second study was conducted with 245 pediatric patients using Seeplex Diarrhea ACE assays (ACE-Bacteria 1, Bacteria 2 and Viral assays) collectively detecting 15 enteric pathogens, including Salmonella spp., Shigella spp., Vibrio spp., toxin B producer Clostridium difficile, Campylobacter spp., Clostridium perfringens, Yersinia enterocolitica, Aeromonas spp., Escherichia coli O157:H7, verocytotoxin-producing Escherichia coli , Adenovirus, group A rotavirus, Norovirus GI and GII, and Astrovirus 73 . This study showed better sensitivity for multiplex PCR than routine methods, except for Salmonella spp.…”

Section: Current Enteric Multiplex Commercial Assaysmentioning

confidence: 99%

Multiplex Polymerase Chain Reaction Tests for Detection of Pathogens Associated with Gastroenteritis

Zhang

Morrison

Tang

2015

Clinics in Laboratory Medicine

147

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Synopsis A wide range of enteric pathogens can cause infectious gastroenteritis. Conventional diagnostic algorithms including culture, biochemical identification, immunoassay and microscopic examination are time consuming and often lack sensitivity and specificity. Advances in molecular technology have as allowed its use as clinical diagnostic tools. Multiplex PCR based testing has made its way to gastroenterology diagnostic arena in recent years. In this article we present a review of recent laboratory developed multiplex PCR tests and current commercial multiplex gastrointestinal pathogen tests. We will focus on two FDA cleared commercial syndromic multiplex tests: Luminex xTAG GPP and Biofire FimArray GI test. These multiplex tests can detect and identify multiple enteric pathogens in one test and provide results within hours. Multiplex PCR tests have shown superior sensitivity to conventional methods for detection of most pathogens. The high negative predictive value of these multiplex tests has led to the suggestion that they be used as screening tools especially in outbreaks. Although the clinical utility and benefit of multiplex PCR test are to be further investigated, implementing these multiplex PCR tests in gastroenterology diagnostic algorithm has the potential to improve diagnosis of infectious gastroenteritis.

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“…(Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus), and Clostridium difficile toxin B. This PCR assay has been previously validated and is reported to be more sensitive than culture for detection of Campylobacter from stool (10,11).…”

mentioning

confidence: 99%

Multicenter Evaluation of Clinical Diagnostic Methods for Detection and Isolation of Campylobacter spp. from Stool

et al. 2016

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The use of culture-independent diagnostic tests (CIDTs), such as stool antigen tests, as standalone tests for the detection of Campylobacter in stool is increasing. We conducted a prospective, multicenter study to evaluate the performance of stool antigen CIDTs compared to culture and PCR for Campylobacter detection. Between July and October 2010, we tested 2,767 stool specimens from patients with gastrointestinal illness with the following methods: four types of Campylobacter selective media, four commercial stool antigen assays, and a commercial PCR assay. Illnesses from which specimens were positive by one or more culture media or at least one CIDT and PCR were designated "cases." A total of 95 specimens (3.4%) met the case definition. The stool antigen CIDTs ranged from 79.6% to 87.6% in sensitivity, 95.9 to 99.5% in specificity, and 41.3 to 84.3% in positive predictive value. Culture alone detected 80/89 (89.9% sensitivity) Campylobacter jejuni/Campylobacter coli-positive cases. Of the 209 noncases that were positive by at least one CIDT, only one (0.48%) was positive by all four stool antigen tests, and 73% were positive by just one stool antigen test. The questionable relevance of unconfirmed positive stool antigen CIDT results was supported by the finding that noncases were less likely than cases to have gastrointestinal symptoms. Thus, while the tests were convenient to use, the sensitivity, specificity, and positive predictive value of Campylobacter stool antigen tests were highly variable. Given the relatively low incidence of Campylobacter disease and the generally poor diagnostic test characteristics, this study calls into question the use of commercially available stool antigen CIDTs as standalone tests for direct detection of Campylobacter in stool. Campylobacter infection continues to be a major public health problem. Campylobacter jejuni and Campylobacter coli are pathogens transmitted commonly through food, causing an estimated 1.3 million cases of illness per year in the United States (1), and yet diagnosis can be challenging because the organism is difficult to isolate, grow, and identify. Recent reports describing clinical laboratory practices for Campylobacter diagnostics in Pennsylvania (2) and the Foodborne Diseases Active Surveillance Network (FoodNet) sites (3) highlight the wide range of testing practices in use; currently, no best-practice clinical or public health laboratory guidelines exist for laboratory diagnosis of Campylobacter infections. Direct plating onto a Campylobacter selective medium, followed by incubation at 42°C under microaerobic conditions for 72 h, has long been considered the "gold standard" for diagnosis (4).The use of culture-independent diagnostic tests (CIDTs) for Campylobacter testing on stool samples is increasing, which may have important implications for both patient management and public health surveillance efforts (5). Stool antigen tests to directly detect Campylobacter in fecal samples are fast and generate sameday results, but concerns regarding speci...

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Evaluation of a multiplex PCR assay for simultaneous detection of bacterial and viral enteropathogens in stool samples of paediatric patients

Cited by 73 publications

References 28 publications

Co-infection as a confounder for the role of Clostridium difficile infection in children with diarrhoea: a summary of the literature

Co-infection as a confounder for the role of Clostridium difficile infection in children with diarrhoea: a summary of the literature

Multiplex Polymerase Chain Reaction Tests for Detection of Pathogens Associated with Gastroenteritis

Multicenter Evaluation of Clinical Diagnostic Methods for Detection and Isolation of Campylobacter spp. from Stool

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