Performance of commercial PCR assays to detect toxigenic <i>Clostridioides difficile</i> in the feces of puppies

Anis, Eman; Barnart, Denise; Barnard, Amanda; Kelly, Douglas G.; Redding, Laurel E.

doi:10.1002/vms3.567

Cited by 5 publications

(4 citation statements)

References 29 publications

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“…difficile that has not been validated in bovids could have resulted in false negatives or false positives. However, we believe this possibility to be minimal, because we have shown that the performance of this assay is relatively high in other species [ 16 ], and because pooled samples underwent anaerobic culture, which would have ruled out false negatives. Finally, because the qPCR assay only detects toxigenic C. difficile, we were unable to detect non-toxigenic C .…”

Section: Discussionmentioning

confidence: 99%

“…Additionally, the assay has a callout for ribotype NAP1/B1/027. While this assay has not been validated for use in bovine feces, the sensitivity and specificity of this assay in human stools are 93.5% (95% CI: 90.3–95.9%) and 94.0% (95% CI: 92.9–95.0%), respectively, according to the manufacturer, and 84.2% and 87.7%, respectively, in canine feces [ 16 ].…”

Section: Methodsmentioning

confidence: 99%

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Gut microbiota features associated with Clostridioides difficile colonization in dairy calves

et al. 2021

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Diarrheal disease, a major cause of morbidity and mortality in dairy calves, is strongly associated with the health and composition of the gut microbiota. Clostridioides difficile is an opportunistic pathogen that proliferates and can produce enterotoxins when the host experiences gut dysbiosis. However, even asymptomatic colonization with C. difficile can be associated with differing degrees of microbiota disruption in a range of species, including people, swine, and dogs. Little is known about the interaction between C. difficile and the gut microbiota in dairy calves. In this study, we sought to define microbial features associated with C. difficile colonization in pre-weaned dairy calves less than 2 weeks of age. We characterized the fecal microbiota of 80 calves from 23 different farms using 16S rRNA sequencing and compared the microbiota of C. difficile-positive (n = 24) and C. difficile-negative calves (n = 56). Farm appeared to be the greatest source of variability in the gut microbiota. When controlling for calf age, diet, and farm location, there was no significant difference in Shannon alpha diversity (P = 0.50) or in weighted UniFrac beta diversity (P = 0.19) between C. difficile-positive and–negative calves. However, there was a significant difference in beta diversity as assessed using Bray-Curtiss diversity (P = 0.0077), and C. difficile-positive calves had significantly increased levels of Ruminococcus (gnavus group) (Adj. P = 0.052), Lachnoclostridium (Adj. P = 0.060), Butyricicoccus (Adj. P = 0.060), and Clostridium sensu stricto 2 compared to C. difficile-negative calves. Additionally, C. difficile-positive calves had fewer microbial co-occurrences than C. difficile–negative calves, indicating reduced bacterial synergies. Thus, while C. difficile colonization alone is not associated with dysbiosis and is therefore unlikely to result in an increased likelihood of diarrhea in dairy calves, it may be associated with a more disrupted microbiota.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Gut microbiota features associated with Clostridioides difficile colonization in dairy calves

et al. 2021

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“…Pet owners from the UPHS cohort were mailed stool collection kits with instructions to collect a faecal sample from their pet(s) (cats or dogs only) within 2 weeks of their positive C. difficile test and ship it back overnight to the laboratory in an insulated package with an ice pack. These methods have been used successfully by our team to obtain C. difficile‐ positive stool samples from people and dogs (Anis et al, 2021; L. Redding et al, 2021).…”

Section: Methodsmentioning

confidence: 99%

“…To confirm culture results for toxigenic isolates, all samples underwent qPCR using an in‐house multiplex PCR that detects the tcdA and tcdB genes and has been validated for use in canine stool (Anis et al, 2021).…”

Section: Methodsmentioning

confidence: 99%

Infrequent intrahousehold transmission of Clostridioides difficile between pet owners and their pets

Redding

Habing

et al. 2023

Zoonoses and Public Health

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Companion animals have been shown to carry Clostridioides difficile strains that are similar or identical to strains found in people, and a small number of studies have shown that pets carry genetically identical C. difficile isolates as their owners, suggesting inter‐species transmission. However, the directionality of transmission is ultimately unknown, and the frequency with which animals acquire C. difficile following their owners' infection is unclear. The goal of this study was to assess how often pets belonging to people with C. difficile infection carry genetically related C. difficile isolates. We enrolled pet owners from two medical institutions (University of Pennsylvania Health System (UPHS) and The Ohio State University Wexner Medical Center (OSUWMC)) who had diarrhoea with or without positive C. difficile assays and tested their faeces and their pets' faeces for C. difficile using both anaerobic culture and PCR assays. When microorganisms were obtained from both the owner and pet and had the same toxin profile or ribotype, isolates underwent genomic sequencing. Faecal samples were obtained from a total of 59 humans, 72 dogs and 9 cats, representing 47 complete households (i.e. where a sample was available from the owner and at least one pet). Of these, C. difficile was detected in 30 humans, 10 dogs and 0 cats. There were only two households where C. difficile was detected in both the owner and pet. In one of these households, the C. difficile isolates were of different toxin profiles/ribotypes (A+/B+ / RT 499 from the owner, A‐/B‐ / RT PR22386 from the dog). In the other household, the isolates were genetically identical (one SNP difference). Interestingly, the dog from this household had recently received a course of antibiotics (cefpodoxime and metronidazole). Our findings suggest that inter‐species transmission of C. difficile occurs infrequently in households with human C. difficile infections.

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Performance of commercial PCR assays to detect toxigenic Clostridioides difficile in the feces of puppies

Anis

Barnart

Barnard

et al. 2021

Veterinary Medicine & Sci

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Clostridioides difficile is an important enteric pathogen that causes significant morbidity and mortality in humans. With community‐acquired infections on the rise, it is important to identify reservoirs of the pathogen. Companion animals can be asymptomatic carriers of C. difficile and may therefore represent a reservoir, but epidemiological studies of C. difficile within the pet‐owner unit are needed, along with validated methods to detect C. difficile in both people and animals. The goal of this study was to assess the performance of commercial qPCR assays and a multiplex PCR for C. difficile compared to toxigenic culture. These assays were tested on up to 103 fecal samples from puppies, a population in which the prevalence of C. difficile is the highest. The sensitivities, specificities, positive predictive values and negative predictive values were respectively 84.2%, 87.7%, 61.5%, and 95.9% for the Cepheid GeneXpert; 66.7%, 66.7%, 29.6%, and 90.9% for the DiaSorin Simplexa; and 94.4%, 85.0%, 65.4%, and 98.1%, for the multiplex qPCR. The agreement was highest between the GeneXpert and the multiplex PCR (90.1% agreement, with a kappa statistic of 0.77). For diagnostic purposes, the positive predictive values of the assays were low. However, the high sensitivities of the assays could render them useful for epidemiologic purposes.

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Performance of commercial PCR assays to detect toxigenic Clostridioides difficile in the feces of puppies

Cited by 5 publications

References 29 publications

Gut microbiota features associated with Clostridioides difficile colonization in dairy calves

Gut microbiota features associated with Clostridioides difficile colonization in dairy calves

Infrequent intrahousehold transmission of Clostridioides difficile between pet owners and their pets

Performance of commercial PCR assays to detect toxigenic Clostridioides difficile in the feces of puppies

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