Verification and large scale clinical evaluation of a national standard protocol for Salmonella spp ./Shigella spp. screening using real-time PCR combined with guided culture

Tang, Xi-Jun; Yang, Zexiao; Chen, Xinbin; Tian, Wen‐de; Tu, Cheng-Ning; Wang, Haibo

doi:10.1016/j.mimet.2017.12.007

Cited by 17 publications

(10 citation statements)

References 24 publications

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“…Boer et al 17 found that the number of Salmonella positive samples increased by 2.2% when qPCR was performed after enrichment in selenite broth. Tang et al 18 have also validated the high sensitivity and specificity of a national standard protocol based on Real-Time PCR combined with guided culture. www.nature.com/scientificreports www.nature.com/scientificreports/ Relative to Campylobacter, the qPCR assays exhibited notable differences in their abilities to detect many Campylobacter species.…”

Section: Discussionmentioning

confidence: 99%

Comparison of several Real-Time PCR Kits versus a Culture-dependent Algorithm to Identify Enteropathogens in Stool Samples

Valledor

Gil-Rodríguez

et al. 2020

Sci Rep

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This study aims to validate the current diagnostic method for the clinical detection of gastroenteritis. We analyzed 400 stool samples to detect three of the most common enteropathogens: Salmonella spp., Campylobacter spp., and Yersinia enterocolitica. All specimens were tested with a routine clinical diagnosis algorithm and with five real-time PCR assays. A total of 98 specimens (24.5%) were positive for enteropathogens. We found 24 samples positive for Salmonella enterica, 71 positive for Campylobacter spp., and 4 positive for Yersinia enterocolitica. All evaluated methods exhibited a good performance in identifying Salmonella and Yersinia enterocolitica, being the highest positive percent agreement (PPA) value of 95.8% and 100%, respectively. The clinical algorithm showed the highest ppA value identifying Salmonella, due to the enrichment in selenite broth. However, the evaluated methods showed notable differences in the identification of Campylobacter species, obtaining a wide range of PPA values: 59.2%-100%. The clinical algorithm showed the lowest PPA value since it was only able to detect Campylobacter jejuni and Campylobacter coli species. This study revealed the importance of implementing the real-time PCR technique in a clinical algorithm: it improved the accuracy of the diagnosis and provided results in a shorter time compared to routine clinical methods. Acute gastroenteritis is a leading cause of morbidity and mortality worldwide 1 , particularly in at-risk populations, such as children, older individuals, and immunocompromised patients. We included three bacteria in this study: Salmonella spp., Campylobacter spp., and Yersinia enterocolitica, which are among the most common enteropathogens that cause gastrointestinal infections. Thus, their rapid, accurate identification is crucial for infection control, and in selected cases, for determining the most suitable therapy. According to the Centers for Disease Control and Prevention, during 2016, the Foodborne Active Surveillance Network identified 24,029 cases of foodborne infections, which led to 5,512 hospitalizations (22.9%) and 98 deaths (0.4%). Some of these cases were detected by culture, but others were detected with culture-independent diagnostic tests (CIDTs). Current CIDTs mainly comprise two types: antigen-based and nucleic acid-based assays. In 2016, one study showed that many infections were identified with a CIDT and without a culture confirmation; most frequently, Campylobacter (32%) and Yersinia (32%); but also Shiga toxin-producing E.coli (STEC) (24%), Shigella (23%), Vibrio (13%), and Salmonella (8%) 2. That study revealed the consequences of the current change in diagnostic trends. The implementation of CIDTs in clinical laboratories has led to more exhaustive diagnoses. Most of these tests are ordered by clinicians, because CIDTs are easier to perform and they return results faster than traditional culture techniques. Moreover, samples are subjected to deep screening when multiplex panels are used; consequently, the number of reporte...

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Section: Discussionmentioning

confidence: 99%

Comparison of several Real-Time PCR Kits versus a Culture-dependent Algorithm to Identify Enteropathogens in Stool Samples

Valledor

Gil-Rodríguez

et al. 2020

Sci Rep

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“…This result is comparable to both our and other previous reports. In our previous study, the LOD for Salmonella and Shigella was 10 3 CFU/mL and 10 1 CFU/mL, respectively [ 6 ]. In other duplex assays, Cunningham et al reported LOD of 990 CFU/mL and 52 CFU/mL, while Van Lint et al reported LOD of 5528 CFU/mL and 61 CFU/mL for Salmonella and Shigella , respectively [ 15 , 16 ].…”

Section: Discussionmentioning

confidence: 99%

“…Human stool samples were collected using a stool tube from food handlers who came to our center for obligatory Salmonella and Shigella screening during 2017–2019 and first diagnosed by the bacterial culture method as described previously [ 6 ]. 183 samples, including 36 samples positive for Salmonella , 28 samples positive for Shigella, and 119 negative samples, were selected randomly.…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, the lower limit of detection (LOD) of the method may be as high as 10 4 CFU/mL [ 6 ]. With the aim of improving the effectiveness and sensitivity, we developed several nucleic acid-based assays, including PCR [ 6 , 7 ], DNA microarray [ 8 ], and real-time nucleic acid sequence-based amplification (RT-NASBA) [ 9 ], for the detection of Salmonella and Shigella or other diarrhea-related pathogens. However, these assays are not suitable for on-site detection, where no sophisticated thermal cycler is available.…”

Section: Introductionmentioning

confidence: 99%

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Development and Evaluation of an iiPCR Assay for Salmonella and Shigella Detection on a Field-Deployable PCR System

Lin

Zhao³

et al. 2020

Canadian Journal of Infectious Diseases and Medical Microbiolog

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Background. Salmonella and Shigella are often associated with fecal-oral transmission and cause large-scale outbreaks in centralized catering units and, therefore, should be frequently and strictly monitored, especially among food handlers. However, no specific and sensitive on-site detection method is available until now. Methods. In this study, an insulated isothermal PCR assay for the detection of Salmonella and Shigella on a field-deployable PCR system was developed. Specificity, sensitivity, reproducibility, and clinical accuracy of the assay were characterized and evaluated. Results. The insulated isothermal PCR assay could be completed within 58 minutes with minimal pretreatment needed. The assay was specific and with good reproducibility. The limit of detection was 103 CFU/mL and 101 CFU/mL for Salmonella and Shigella, respectively, which was comparable to multiplex real-time PCR. Mock on-site clinical evaluation results showed that the analytical sensitivity and specificity of the insulated isothermal PCR assay were 100% and 96.6%, while the positive predictive value and negative predictive value were 94.1% and 100%, respectively. Conclusion. Based on our results, we believe that the assay developed herein could serve as an alternative method for preliminary screening and provide a valuable platform for the on-site detection of Salmonella and Shigella, especially in resource-limited and developing countries.

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“…Multiplex polymerase chain reaction (PCR), technique is also available commercially (Kimberlin et al 2015 ). The culture-based approaches inherently possess low sensitivity, as the resistance shown by S. sonnei to ciprofloxacin, erythromycin and azithromycin has become a serious problem (Tang et al 2018 ).…”

Section: Introductionmentioning

confidence: 99%

Shigella sonnei: virulence and antibiotic resistance

Shad

2020

Arch Microbiol

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Shigella sonnei is the emerging pathogen globally, as it is the second common infectious species of shigellosis (bloody diarrhoea) in low-and middle-income countries (LMICs) and the leading one in developed world. The multifactorial processes and novel mechanisms have been identified in S. sonnei, that are collectively playing apart a substantial role in increasing its prevalence, while replacing the S. flexneri and other Gram-negative gut pathogens niche occupancy. Recently, studies suggest that due to improvement in sanitation S. sonnei has reduced cross-immunization from Plesiomonas shigelliodes (having same O-antigen as S. sonnei) and also found to outcompete the two major species of Enterobacteriaceae family (Shigella flexneri and Escherichia coli), due to encoding of type VI secretion system (T6SS). This review aimed to highlight S. sonnei as an emerging pathogen in the light of recent research with pondering aspects on its epidemiology, transmission, and pathogenic mechanisms. Additionally, this paper aimed to review S. sonnei disease pattern and related complications, symptoms, and laboratory diagnostic techniques. Furthermore, the available treatment reigns and antibiotic-resistance patterns of S. sonnei are also discussed, as the ciprofloxacin and fluoroquinolone-resistant S. sonnei has already intensified the global spread and burden of antimicrobial resistance. In last, prevention and controlling strategies are briefed to limit and tackle S. sonnei and possible future areas are also explored that needed more research to unravel the hidden mysteries surrounding S. sonnei.

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Verification and large scale clinical evaluation of a national standard protocol for Salmonella spp ./Shigella spp. screening using real-time PCR combined with guided culture

Cited by 17 publications

References 24 publications

Comparison of several Real-Time PCR Kits versus a Culture-dependent Algorithm to Identify Enteropathogens in Stool Samples

Comparison of several Real-Time PCR Kits versus a Culture-dependent Algorithm to Identify Enteropathogens in Stool Samples

Development and Evaluation of an iiPCR Assay for Salmonella and Shigella Detection on a Field-Deployable PCR System

Shigella sonnei: virulence and antibiotic resistance

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