A novel multiplex PCR assay for<i>Salmonella</i>subspecies identification

Lee, Ken-ichi; Iwata, Tadahisa; Shimizu, Manabu; Taniguchi, Takahide; Nakadai, Aya; Hirota, Yoshikazu; Hayashidani, Hideki

doi:10.1111/j.1365-2672.2009.04263.x

Cited by 43 publications

(33 citation statements)

References 31 publications

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“…This could limit serotype exposure and allow cross contamination between individuals through husbandry. The Salmonella serotype II 13,22:z 29 :1,5 was found in 4 green iguanas in the current study, has previously been described in a pet turtle in Japan and free living tortoises in Morocco (Hidalgo-Vila et al 2008;Lee et al 2009). Serotype IIIb 6,14:z10:z was found in 4 green iguanas and has been reported in captive snakes in the United States (Goupil et al 2012).…”

Section: Discussionsupporting

confidence: 77%

Choanal and cloacal aerobic bacterial flora in captive green iguanas: a comparative analysis

et al. 2015

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The aims of this study were to characterize the choanal and cloacal aerobic bacterial flora in healthy captive green iguanas and to compare it with the bacterial flora of the biofilm present in the water container of each terrarium. Samples were collected from the choana and the cloaca of 20 healthy captive adult green iguanas and from the biofilm of 15 water containers. The final identification of aerobic bacteria was performed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Salmonella positive samples were serotyped. The most common strains observed at each test location were from 1) choanae: Staphylococcus spp., Enterobacter cloacae and Comamonas testosteroni; 2) cloacae: Citrobacter spp., Salmonella spp. and Corynebacterium spp.; and 3) biofilms: Pseudomonas spp., Salmonella spp. and Acidovorax spp. We showed that apart from Salmonella spp., the choanal and cloacal bacterial flora differed from the microorganisms present in the biofilm of the animal's water container. These data revealed that healthy captive adult green iguanas harbored several aerobic bacterial strains that in immunosuppressed reptiles may act as opportunistic pathogens. Also, several of the aerobic bacteria identified in samples are potential zoonotic agents. Characterization of the normal background flora in captive reptiles and their environment can contribute to an understanding of the spread of bacterial contamination and the risk of potential zoonotic diseases for people in contact with these animals. Reptiles, biofilm, matrix-assisted laser desorption ionization time-of-flight mass spectrometry, serotypePrevious studies in clinically healthy captive lizards showed a predominance of Pseudomonas spp., Aeromonas spp., Serratia spp. and Enterobacter spp. in the oral cavity of green iguanas (Barten 2002) and Citrobacter spp., Enterobacter spp., Escherichia coli, Klebsiella oxytoca, Salmonella spp., Pseudomonas aeruginosa, Corynebacterium spp., Staphylococcus spp., and Streptococcus spp. in the cloaca of giant lizards (Martínez Silvestre et al. 2003). These studies have helped us to understand the normal bacterial flora in captive lizards and to interpret the significance of bacterial strains in ill individuals.Pet green iguanas are kept for many years in indoor terrariums. This situation leads us to consider the influence of the bacterial population in the biofilm that forms on the water container that is used by the lizard to drink from and bathe in, on the bacterial community of the lizard.The aims of this study were to characterize and compare the aerobic bacterial flora between the choanal and cloacal sites in healthy captive adult green iguanas, and with the bacteria of the water container in their terrariums. This information will explore the hypothesis that the biofilm in water containers may perpetuate and spread bacterial contamination between animals and between terrariums. Furthermore, these data will contribute to knowledge of the normal background flora in pet iguanas and their environ...

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Section: Discussionsupporting

confidence: 77%

Choanal and cloacal aerobic bacterial flora in captive green iguanas: a comparative analysis

et al. 2015

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“…Although the cell lysates were not included in the final protocol, the results showed promise, and since several groups have demonstrated the success of optimization of multiplex PCRs for use in Salmonella molecular typing methods, the use of lysates in investigations of future layouts is planned (7,22,23). A study describing a multiplex PCR assay designed to detect all of the Salmonella subspecies, including the species S. bongori, was recently reported (33). Adaptation of this method is planned to be investigated as an additional multiplex reaction in the SGSA protocol, and subspecies-specific probes are to be added to the array for verification of all six distinct subspecies.…”

Section: Discussionmentioning

confidence: 99%

Rapid Genoserotyping Tool for Classification of Salmonella Serovars

et al. 2011

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We have developed a Salmonella genoserotyping array (SGSA) which rapidly generates an antigenic formula consistent with the White-Kauffmann-Le Minor scheme, currently the gold standard for Salmonella serotyping. A set of 287 strains representative of 133 Salmonella serovars was assembled to validate the array and to test the array probes for accuracy, specificity, and reproducibility. Initially, 76 known serovars were utilized to validate the specificity and repeatability of the array probes and their expected probe patterns. The SGSA generated the correct serovar designations for 100% of the known subspecies I serovars tested in the validation panel and an antigenic formula consistent with that of the White-Kauffmann-Le Minor scheme for 97% of all known serovars tested. Once validated, the SGSA was assessed against a blind panel of 100 Salmonella enterica subsp. I samples serotyped using traditional methods. In summary, the SGSA correctly identified all of the blind samples as representing Salmonella and successfully identified 92% of the antigens found within the unknown samples. Antigen-and serovar-specific probes, in combination with a pepT PCR for confirmation of S. enterica subsp. Enteritidis determinations, generated an antigenic formula and/or a serovar designation consistent with the White-Kauffmann-Le Minor scheme for 87% of unknown samples tested with the SGSA. Future experiments are planned to test the specificity of the array probes with other Salmonella serovars to demonstrate the versatility and utility of this array as a public health tool in the identification of Salmonella.

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“…The following cycling conditions were performed: initial denaturation at 94 °C for 5 min, followed by 30 cycles of 55 °C for 30 s, 72 °C for 1 min and 94 °C for 30 s, and a final extension at 72 °C for 3 min. Samples which were 288 d -differs among strains positive for Salmonella were then examined by multiplex PCR amplification based on formula described previously by Lee et al (2009). The list of primers used in multiplex PCR which give unique results for each subspecies of Salmonella strains are shown in Table 1.…”

Section: Methodsmentioning

confidence: 99%

Salmonella serovar spectrum associated with reptiles in Poland

2014

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This study aimed to evaluate the incidence of Salmonella isolates from a wide variety of reptiles in Poland. A total of 374 faecal samples from chelonians, lizards and snakes were collected between 2009 and 2012. The nested, two-step PCR and multiplex PCR were performed to access the incidence and to characterize Salmonella isolates. Salmonella strains were found in 122 of 374 samples (32.6%). Among the different reptilian species, Salmonella strains were found in 58 samples from lizards (38.9%), 31 samples from snakes (28.7%) and 33 samples from chelonians (28.2%). Of the total of 122 strains, 72 belonged to the species Salmonella enterica subsp. enterica, 20 to the species S. enterica subs. salamae or S. enterica subs. houtanae. The incidence of S. enterica subs. diarizonae and S. enterica subs. indica was low, constituting less than 3.5% of the examined population. The findings show that reptiles can be considered as a reservoir for Salmonella and hence could pose a zoonotic hazard. In addition, multiplex PCR assay is a rapid, specific and easy-to-perform method and might be applied for rapid screening of large numbers of Salmonella samples.

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A novel multiplex PCR assay forSalmonellasubspecies identification

Cited by 43 publications

References 31 publications

Choanal and cloacal aerobic bacterial flora in captive green iguanas: a comparative analysis

Choanal and cloacal aerobic bacterial flora in captive green iguanas: a comparative analysis

Rapid Genoserotyping Tool for Classification of Salmonella Serovars

Salmonella serovar spectrum associated with reptiles in Poland

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