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Transcriptome analysis of the biofilm formed by methicillin-susceptible Staphylococcus aureus.

Tan X, Qin N, Wu C, Sheng J, Yang R, Zheng B, Ma Z, Liu L, Peng X, Jia A - Sci Rep (2015)

Bottom Line: These findings suggest that biofilms of S. aureus with agr dysfunction may be more resistant than those with agr function.Therefore, the infection from clinical MSSA may be recalcitrant once forming biofilm.Further study is necessary to uncover the mechanisms of biofilm formation in other clinical S. aureus.

View Article: PubMed Central - PubMed

Affiliation: School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, China.

ABSTRACT
Biofilm formation is regarded as one of the major determinants in the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) as pathogens of medical device-related infection. However, methicillin-susceptible S. aureus (MSSA) can also form biofilm in vitro and such biofilms are resistant to vancomycin. Hence, researching the possible mechanisms of MSSA biofilm formation is urgent and necessary. Here, we used S. aureus ATCC25923 as the model strain, and studied gene expression profiles in biofilms after the treatment of ursolic acid and resveratrol using RNA-seq technology. The results showed that only ursolic acid could inhibit biofilm formation, which differed from their applied on the multiple clinical drugs resistant MRSA biofilm. RNA-seq data was validated by examining the expression of six genes involved in biofilm formation by qRT-PCR. These data analysis indicated that the mechanism of the MSSA biofilm formation was different from that of the MRSA, due to absence of accessory gene regulator (agr) function. These findings suggest that biofilms of S. aureus with agr dysfunction may be more resistant than those with agr function. Therefore, the infection from clinical MSSA may be recalcitrant once forming biofilm. Further study is necessary to uncover the mechanisms of biofilm formation in other clinical S. aureus.

No MeSH data available.


Related in: MedlinePlus

PCR product agarose gel electrophoresis for the identification of S. aureus ATCC25923 agr genotype.Whole- cell PCR was performed with each of the four agr specificity primers. Lines 1 to 4, PCR product using the same forward primer pan-agr and different reverse primers (agrI, agrII, agrIII, and agrIV) with S. aureus ATCC25923 genomic DNA as templates (439 bp for agrI, 573 bp for agrII, 406 bp for agrIII, and 657 bp for agrIV); Lines 5 to 8, negative controls responsible to lines 1 to 4, respectively; Lines M, 100 bp for DNA ladder.
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f3: PCR product agarose gel electrophoresis for the identification of S. aureus ATCC25923 agr genotype.Whole- cell PCR was performed with each of the four agr specificity primers. Lines 1 to 4, PCR product using the same forward primer pan-agr and different reverse primers (agrI, agrII, agrIII, and agrIV) with S. aureus ATCC25923 genomic DNA as templates (439 bp for agrI, 573 bp for agrII, 406 bp for agrIII, and 657 bp for agrIV); Lines 5 to 8, negative controls responsible to lines 1 to 4, respectively; Lines M, 100 bp for DNA ladder.

Mentions: S. aureus ATCC25923 agr specificity was identified by the expected PCR product size according to Shopsin et al. report21. The lengths of the PCR products were estimated by comparing with the 100 bp DNA ladder (Takara, Japan). The results showed that only agr-III genotype was obtained by the PCR assay and the length of the PCR product was 406 bp, which corresponded to expected product size (Fig. 3). In addition, we also sequenced the PCR product and aligned it with Staphylococcus aureus RN8462 (GenBank accession number AF001783) using BLAST, which is agr group III strain102223. The results revealed that their identity was 99% and amplified fragment corresponding to the region was right. Therefore, based on above results, we determined that S. aureus ATCC25923 was agr-III genotype strain.


Transcriptome analysis of the biofilm formed by methicillin-susceptible Staphylococcus aureus.

Tan X, Qin N, Wu C, Sheng J, Yang R, Zheng B, Ma Z, Liu L, Peng X, Jia A - Sci Rep (2015)

PCR product agarose gel electrophoresis for the identification of S. aureus ATCC25923 agr genotype.Whole- cell PCR was performed with each of the four agr specificity primers. Lines 1 to 4, PCR product using the same forward primer pan-agr and different reverse primers (agrI, agrII, agrIII, and agrIV) with S. aureus ATCC25923 genomic DNA as templates (439 bp for agrI, 573 bp for agrII, 406 bp for agrIII, and 657 bp for agrIV); Lines 5 to 8, negative controls responsible to lines 1 to 4, respectively; Lines M, 100 bp for DNA ladder.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493712&req=5

f3: PCR product agarose gel electrophoresis for the identification of S. aureus ATCC25923 agr genotype.Whole- cell PCR was performed with each of the four agr specificity primers. Lines 1 to 4, PCR product using the same forward primer pan-agr and different reverse primers (agrI, agrII, agrIII, and agrIV) with S. aureus ATCC25923 genomic DNA as templates (439 bp for agrI, 573 bp for agrII, 406 bp for agrIII, and 657 bp for agrIV); Lines 5 to 8, negative controls responsible to lines 1 to 4, respectively; Lines M, 100 bp for DNA ladder.
Mentions: S. aureus ATCC25923 agr specificity was identified by the expected PCR product size according to Shopsin et al. report21. The lengths of the PCR products were estimated by comparing with the 100 bp DNA ladder (Takara, Japan). The results showed that only agr-III genotype was obtained by the PCR assay and the length of the PCR product was 406 bp, which corresponded to expected product size (Fig. 3). In addition, we also sequenced the PCR product and aligned it with Staphylococcus aureus RN8462 (GenBank accession number AF001783) using BLAST, which is agr group III strain102223. The results revealed that their identity was 99% and amplified fragment corresponding to the region was right. Therefore, based on above results, we determined that S. aureus ATCC25923 was agr-III genotype strain.

Bottom Line: These findings suggest that biofilms of S. aureus with agr dysfunction may be more resistant than those with agr function.Therefore, the infection from clinical MSSA may be recalcitrant once forming biofilm.Further study is necessary to uncover the mechanisms of biofilm formation in other clinical S. aureus.

View Article: PubMed Central - PubMed

Affiliation: School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, China.

ABSTRACT
Biofilm formation is regarded as one of the major determinants in the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) as pathogens of medical device-related infection. However, methicillin-susceptible S. aureus (MSSA) can also form biofilm in vitro and such biofilms are resistant to vancomycin. Hence, researching the possible mechanisms of MSSA biofilm formation is urgent and necessary. Here, we used S. aureus ATCC25923 as the model strain, and studied gene expression profiles in biofilms after the treatment of ursolic acid and resveratrol using RNA-seq technology. The results showed that only ursolic acid could inhibit biofilm formation, which differed from their applied on the multiple clinical drugs resistant MRSA biofilm. RNA-seq data was validated by examining the expression of six genes involved in biofilm formation by qRT-PCR. These data analysis indicated that the mechanism of the MSSA biofilm formation was different from that of the MRSA, due to absence of accessory gene regulator (agr) function. These findings suggest that biofilms of S. aureus with agr dysfunction may be more resistant than those with agr function. Therefore, the infection from clinical MSSA may be recalcitrant once forming biofilm. Further study is necessary to uncover the mechanisms of biofilm formation in other clinical S. aureus.

No MeSH data available.


Related in: MedlinePlus