Rectal swab sampling followed by an enrichment culture-based real-time PCR assay to detect Salmonella enterocolitis in children

Lin, Lung–Huang; Tsai, Chieh-Yuan; Hung, Min-Hsuan; Fang, Ya-Chieh; Ling, Qing-Dong

doi:10.1111/j.1469-0691.2010.03450.x

Cited by 12 publications

(10 citation statements)

References 22 publications

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“…Hence, molecular approaches for Salmonella identification characterized by high sensitivity and a short turnaround time have been developed. PCR-based assays have been clinically validated for diagnosis of gastrointestinal NTS infection (118), invasive NTS infection (119), and typhoidal Salmonella infection in the blood of patients with enteric fever (reviewed in reference 120) and are increasingly being used for diagnosis of Salmonella infections. Lin and associates successfully demonstrated the feasibility of a bacterial enrichment culture-based real-time PCR for detection and screening of NTS infection in children presenting with diarrhea.…”

Section: Pcr-based Molecular Methods For Salmonella Detectionmentioning

confidence: 99%

Persistent Infection and Long-Term Carriage of Typhoidal and Nontyphoidal Salmonellae

2018

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SUMMARY The ability of pathogenic bacteria to affect higher organisms and cause disease is one of the most dramatic properties of microorganisms. Some pathogens can establish transient colonization only, but others are capable of infecting their host for many years or even for a lifetime. Long-term infection is called persistence, and this phenotype is fundamental for the biology of important human pathogens, including Helicobacter pylori, Mycobacterium tuberculosis, and Salmonella enterica. Both typhoidal and nontyphoidal serovars of the species Salmonella enterica can cause persistent infection in humans; however, as these two Salmonella groups cause clinically distinct diseases, the characteristics of their persistent infections in humans differ significantly. Here, following a general summary of Salmonella pathogenicity, host specificity, epidemiology, and laboratory diagnosis, I review the current knowledge about Salmonella persistence and discuss the relevant epidemiology of persistence (including carrier rate, duration of shedding, and host and pathogen risk factors), the host response to Salmonella persistence, Salmonella genes involved in this lifestyle, as well as genetic and phenotypic changes acquired during prolonged infection within the host. Additionally, I highlight differences between the persistence of typhoidal and nontyphoidal Salmonella strains in humans and summarize the current gaps and limitations in our understanding, diagnosis, and curing of persistent Salmonella infections.

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Section: Pcr-based Molecular Methods For Salmonella Detectionmentioning

confidence: 99%

Persistent Infection and Long-Term Carriage of Typhoidal and Nontyphoidal Salmonellae

2018

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“…PCR assays have been clinically validated for diagnosis of enterocolitis [71], or for use on cultured isolates [72], but not for direct diagnosis of invasive infections.…”

Section: Available Diagnostic Approaches For Intsmentioning

confidence: 99%

Diagnostics for invasive Salmonella infections: Current challenges and future directions

Andrews

Ryan

2015

Vaccine

122

136

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Invasive Salmonellosis caused by Salmonella enterica serotype Typhi or Paratyphi A, B, C, or invasive non-typhoidal Salmonella serotypes, is an immensely important disease cluster for which reliable, rapid diagnostic tests are not available. Blood culture remains the gold standard but is insensitive, slow, and resource-intensive. Existing molecular diagnostics have poor sensitivity due to the low organism burden in bodily fluids. Commercially available serologic tests for typhoidal Salmonella have had limited sensitivity and specificity. In high burden, resource-limited settings, reliance on clinical diagnosis or inaccurate tests often results in frequent, unnecessary treatment, which contributes selective pressure for the emergence of antimicrobial resistance. This practice also results in inadequate therapy for other etiologies of acute febrile illnesses, including leptospirosis and rickettsial infections. A number of novel serologic, molecular, transcriptomic and metabolomic approaches to diagnostics are under development. Target product profiles that outline specific needs may focus development and investment, and establish benchmarks for accuracy, cost, speed, and portability of new diagnostics. Of note, a critical barrier to diagnostic assay rollout will be the low cost and low perceived harm of empiric therapy on behalf of providers and patients, which leaves few perceived incentives to utilize diagnostics. Approaches that align incentives with societal goals of limiting inappropriate antimicrobial use, such as subsidizing diagnostics, may be essential for stimulating development and uptake of such assays in resource-limited settings. New diagnostics for invasive Salmonellosis should be developed and deployed alongside diagnostics for alternative etiologies of acute febrile illnesses to improve targeted use of antibiotics.

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“…Although stool samples are considered the "gold standard" specimen for studying gut bacteria in general (5,6) and for detecting CRE in particular (7), hospital epidemiologists, clinical microbiologists, and researchers frequently use rectal swabs due to practical considerations, such as ease of collection, handling, and processing (8)(9)(10)(11). Several studies compared the qualitative sensitivity of rectal swabs versus stool specimens for the recovery and detection of various pathogens, such as Campylobacter fetus subsp.…”

mentioning

confidence: 99%

Rectal Swabs Are Suitable for Quantifying the Carriage Load of KPC-Producing Carbapenem-Resistant Enterobacteriaceae

Lerner

Romano

Chmelnitsky

et al. 2013

Antimicrob Agents Chemother

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It is more convenient and practical to collect rectal swabs than stool specimens to study carriage of colon pathogens. In this study, we examined the ability to use rectal swabs rather than stool specimens to quantify Klebsiella pneumoniae carbapenemase (KPC)-producing carbapenem-resistant Enterobacteriaceae (CRE). We used a quantitative real-time PCR (qPCR) assay to determine the concentration of the bla KPC gene relative to the concentration of 16S rRNA genes and a quantitative culture-based method to quantify CRE relative to total aerobic bacteria. Our results demonstrated that rectal swabs are suitable for quantifying the concentration of KPC-producing CRE and that qPCR showed higher correlation between rectal swabs and stool specimens than the culture-based method. Klebsiella pneumoniae carbapenemase (KPC)-type enzymes are ␤-lactamases, capable of hydrolyzing all known ␤-lactam antibiotics (1). The spread of the bla KPC genes has led to the emergence of carbapenem-resistant Enterobacteriaceae (CRE), mostly K. pneumoniae, as important nosocomial pathogens causing outbreaks in various parts of the world (2). Infections by these pathogens are severe, with an estimated case fatality rate of 35%, and thus considered a clinical and public health threat (3).Active surveillance of high-risk patients has been advocated in areas where CRE are endemic or where there are ongoing outbreaks (4). Although stool samples are considered the "gold standard" specimen for studying gut bacteria in general (5, 6) and for detecting CRE in particular (7), hospital epidemiologists, clinical microbiologists, and researchers frequently use rectal swabs due to practical considerations, such as ease of collection, handling, and processing (8-11). Several studies compared the qualitative sensitivity of rectal swabs versus stool specimens for the recovery and detection of various pathogens, such as Campylobacter fetus subsp. jejuni, vancomycin-resistant Enterococcus faecium (VRE), and Lawsonia intracellularis, by culturing and/or molecular quantification methods (12)(13)(14). These studies had mixed results, with some studies reporting comparable test sensitivity with either sample type (12-14) and others reporting significantly lower sensitivity with rectal swabs (15, 16). For quantitative testing, rectal swabs are believed to be inadequate due to the high variation in the quantity of fecal material on each swab and the difficulties in determining this quantity (17)(18)(19)(20).As the human colon flora is a diverse ecosystem, which is populated by both anaerobic and aerobic microorganisms (21), development of quantitative methods, such as culture-and quantitative real-time PCR (qPCR)-based techniques is important for studying the bacterial composition of this complex ecosystem. The use of these methods can lead to a thorough knowledge about gastrointestinal community composition, the effects of antibiotics, and their roles in health and disease.Here, we suggest an alternate methodology for quantifying pathogens that would overcome the pr...

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Rectal swab sampling followed by an enrichment culture-based real-time PCR assay to detect Salmonella enterocolitis in children

Cited by 12 publications

References 22 publications

Persistent Infection and Long-Term Carriage of Typhoidal and Nontyphoidal Salmonellae

Persistent Infection and Long-Term Carriage of Typhoidal and Nontyphoidal Salmonellae

Diagnostics for invasive Salmonella infections: Current challenges and future directions

Rectal Swabs Are Suitable for Quantifying the Carriage Load of KPC-Producing Carbapenem-Resistant Enterobacteriaceae

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