Identification by real‐time <scp>PCR</scp> with <scp>SYBR</scp> Green of <i>Leishmania</i> spp. and <i>Serratia marcescens</i> in canine ‘sterile’ cutaneous nodular lesions

Cornegliani, L.; Corona, A.; Vercelli, A.; Roccabianca, P.

doi:10.1111/vde.12208

Cited by 10 publications

(12 citation statements)

References 35 publications

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“…RT-qPCR methods are known to be fast, reliable, and highly e cient. Several single RT-qPCR methods for detecting S. nematodiphila (Hurst et al 2008), S. marcescens (Iwaya et al 2005;Joyner et al 2014;Cornegliani et al 2015) and Salmonella spp. (Perelle et al 2004;Nam et al 2005;Tomás et al 2009) have been described.…”

Section: Introductionmentioning

confidence: 99%

Establishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmealestablishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmeal

Ruan¹,

Wang²,

Zhang³

et al. 2020

Preprint

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Salmonella spp. is a high-risk bacterial pathogen that is monitored in imported animal-derived feedstuffs. Serratiafonticola is the bacterial species most frequently confused with Salmonella spp. in traditional identification methods based on biochemical characteristics, which are time-consuming and labor-intensive, and thus unsuitable for daily inspection and quarantine work. In this study, we established a duplex real-time qPCR method with invA- and gyrB-specific primers and probes corresponding to Salmonella spp. and S. fonticola. The method could simultaneously detect both pathogens in imported feedstuffs, with a minimum limit of detection for Salmonella spp. and S. fonticola of 197 copies/SL and 145 copies/SL, respectively (correlation coefficient R2 = 0.999 in both cases). The amplification efficiency for Salmonella spp. and S. fonticola was 98.346% and 96.49%, respectively. Detection of clinical samples was consistent with method GB/T 13091-2018, and all seven artificially contaminated imported feed samples were positively identified. Thus, the developed duplex real-time qPCR assay displays high specificity and sensitivity, and can be used for the rapid and accurate detection of genomic DNA from Salmonella spp. and S. fonticola within hours. This represents a significant improvement in the efficiency of detection of both pathogens in imported feedstuffs.

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Section: Introductionmentioning

confidence: 99%

Establishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmealestablishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmeal

Ruan¹,

Wang²,

Zhang³

et al. 2020

Preprint

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“…Moreover, depending on their abundance and composition, the presence of a dermal or subcutaneous microbiota could also be clinically relevant [11,16]. In animals with cutaneous lesions, skin biopsies are commonly investigated with cultures and molecular techniques to detect and identify microorganisms that could have an etiologic role [17][18][19][20]. An initial assumption in these investigations is that the dermis and subcutaneous tissues of healthy animals are sterile.…”

Section: Introductionmentioning

confidence: 99%

The microbiota of the surface, dermis and subcutaneous tissue of dog skin

et al. 2020

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Background Studies using highly sensitive molecular techniques have detected bacterial communities below the human epidermis. Depending on their abundance and composition, this finding could be clinically relevant. The aim of this study was to determine if bacteria can be detected in the dermis and subcutaneous tissue of dogs without cutaneous disease using two different approaches: traditional cultures and DNA sequencing of the V4 region of bacterial 16S rRNA gene using next-generation sequencing (NGS). Results Seven healthy dogs were included in the study, and two sets of samples were collected from each subject. Sample sets were composed of a 6-mm abdominal skin biopsy, including epidermis, dermis, and subcutis, a skin surface swab, and an environmental blank sample for contamination control. One set of samples from each dog was submitted for bacterial culture and the other one for bacterial DNA amplification and sequencing. Five different bacterial genera (Staphylococcus, Bacillus, Corynebacterium, Streptococcus, and Enterococcus) were isolated in five out of the seven skin surface swab samples with aerobic microbiological culture methods, while no growth was obtained from the other two samples. Although some DNA could be amplified from epidermal, dermal, and subcutaneous tissue samples, the results of the NGS were similar to those of the blanks. Conclusion When investigated with aerobic microbiological culture methods, the dermis and subcutaneous tissue of dogs are sterile. NGS techniques lead to the detection of some bacterial DNA, similar to the signal detected in blanks, which does not support the presence of a microbiota in dermis or subcutaneous tissue.

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“…Moreover, depending on their abundance and composition, the presence of a dermal or subcutaneous microbiota could also be clinically relevant [14,15]. In animals with cutaneous lesions, skin biopsies are commonly investigated with cultures and molecular techniques to detect and identify microorganisms that could have an etiologic role [16][17][18][19]. An initial assumption in these investigations is that the dermis and subcutaneous tissues of healthy animals are sterile.…”

Section: Introductionmentioning

confidence: 99%

Skin microbiome in healthy dogs: from the surface to the inner layers

García-Fonticoba

Ferrer

Francino

et al. 2020

Preprint

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Background. Studies using highly sensitive molecular techniques have detected bacterial communities below the human epidermis. Depending on their abundance and composition, this finding could be clinically relevant. This possibility, however, has not been investigated in the dog so far. The aim of this study was to determine if bacteria can be detected in the dermis and subcutaneous tissue of healthy dogs using two different approaches: traditional cultures and massive DNA sequencing of the V4 region of bacterial 16S rRNA gene using next-generation sequencing (NGS).Results. Seven healthy dogs were included in the study, and two sets of samples were collected from each subject. Sample sets were composed of a 6-mm abdominal skin biopsy, including epidermis, dermis, and subcutis, a skin surface swab, and an environmental blank sample for contamination control. One set of samples from each dog was submitted for bacterial culture and the other one for bacterial DNA amplification and sequencing. Five different bacterial genera (Staphylococcus, Bacillus, Corynebacterium, Streptococcus, and Enterococcus) were isolated in five out of the seven skin surface swab samples with traditional culture methods, while no growth was obtained from the other two samples. Although some DNA could be amplified from epidermal, dermal, and subcutaneous tissue samples, the results of the NGS were similar to those of the blanks.Conclusion. When investigated with traditional culture methods, the dermis and subcutaneous tissue of healthy dogs are sterile. NGS techniques lead to the detection of some bacterial DNA which, although similar to the signal detected in blanks, does not support the presence of a microbiome in dermis or subcutaneous tissue.

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Identification by real‐time PCR with SYBR Green of Leishmania spp. and Serratia marcescens in canine ‘sterile’ cutaneous nodular lesions

Cited by 10 publications

References 35 publications

Establishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmealestablishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmeal

Establishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmealestablishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmeal

The microbiota of the surface, dermis and subcutaneous tissue of dog skin

Skin microbiome in healthy dogs: from the surface to the inner layers

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