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Application of the Random Amplified Polymorphic DNA (RAPD) Fingerprinting to Analyze Genetic Variation in Community Associated-Methicillin Resistant Staphylococcus Aureus (CA-MRSA) Isolates in Iran

View Article: PubMed Central - PubMed

ABSTRACT

The aim of this study was to apply RAPD technique to analyze the genetic variability among the Iranian CA-MRSA isolates.

The RAPD amplification was implemented on 25 strains isolated from the anterior nares of 410 healthy children using four randomly selected oligonucleotide primers from the stocks available in our laboratory, including the primers 1254, GE6, OLP6 and OLP13 from our stock. The amplified PCR products were detected on a 1.5% agarose gel and subjected to further analysis to establish the band profiles and genetic relationships using the Gel Compar® program.

The Iranian CA-MRSA isolates produced distinct RAPD patterns which varied based on the primer used, however, the primer 1254 revealed highly polymorphic patterns consisting 5 discernable RAPD types (RT), “RT1” (12, 48%), “RT2” (8, 32%), “RT3” (3, 12%), and “RT4 and RT5”, (a single RAPD type each, 4%). Phylogenetic analysis based on RAPD profiles divided most of the CA-MRSA isolates into 2 distinct but related RAPD clusters, a small group and two single unrelated RAPD types.

This study shows that the simple and cost-effective but rather difficult to optimize RAPD fingerprinting could be used to evaluate genetic and epidemiological relationships of CA-MRSA isolates on condition that the patterns are obtained from carefully optimized laboratory tests.

No MeSH data available.


Dendrogram of the nasal carriage CA-MRSA RAPD-PCR DNA fingerprints with the primer 1254; Similarity coefficients are included in the top bar
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Figure 3: Dendrogram of the nasal carriage CA-MRSA RAPD-PCR DNA fingerprints with the primer 1254; Similarity coefficients are included in the top bar

Mentions: Dendrogram analysis of RAPD-PCR amplification patterns of the Iranian isolates of CA-MRSA resulted in formation of 2 distinct but related RAPD clusters. RAPD cluster 1 had the highest similarity (89.9%) with RAPD type 4. The remaining 3 isolates occurred in a small group that showed a similarity of 84.5± 4.6% with the other two clusters and RAPD type 4. The RT 5 also formed a rare single line of descent which was distantly related to other isolates with a similarity of 80.2 ± 4.3 % (Figure 3).


Application of the Random Amplified Polymorphic DNA (RAPD) Fingerprinting to Analyze Genetic Variation in Community Associated-Methicillin Resistant Staphylococcus Aureus (CA-MRSA) Isolates in Iran
Dendrogram of the nasal carriage CA-MRSA RAPD-PCR DNA fingerprints with the primer 1254; Similarity coefficients are included in the top bar
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5016346&req=5

Figure 3: Dendrogram of the nasal carriage CA-MRSA RAPD-PCR DNA fingerprints with the primer 1254; Similarity coefficients are included in the top bar
Mentions: Dendrogram analysis of RAPD-PCR amplification patterns of the Iranian isolates of CA-MRSA resulted in formation of 2 distinct but related RAPD clusters. RAPD cluster 1 had the highest similarity (89.9%) with RAPD type 4. The remaining 3 isolates occurred in a small group that showed a similarity of 84.5± 4.6% with the other two clusters and RAPD type 4. The RT 5 also formed a rare single line of descent which was distantly related to other isolates with a similarity of 80.2 ± 4.3 % (Figure 3).

View Article: PubMed Central - PubMed

ABSTRACT

The aim of this study was to apply RAPD technique to analyze the genetic variability among the Iranian CA-MRSA isolates.

The RAPD amplification was implemented on 25 strains isolated from the anterior nares of 410 healthy children using four randomly selected oligonucleotide primers from the stocks available in our laboratory, including the primers 1254, GE6, OLP6 and OLP13 from our stock. The amplified PCR products were detected on a 1.5% agarose gel and subjected to further analysis to establish the band profiles and genetic relationships using the Gel Compar® program.

The Iranian CA-MRSA isolates produced distinct RAPD patterns which varied based on the primer used, however, the primer 1254 revealed highly polymorphic patterns consisting 5 discernable RAPD types (RT), “RT1” (12, 48%), “RT2” (8, 32%), “RT3” (3, 12%), and “RT4 and RT5”, (a single RAPD type each, 4%). Phylogenetic analysis based on RAPD profiles divided most of the CA-MRSA isolates into 2 distinct but related RAPD clusters, a small group and two single unrelated RAPD types.

This study shows that the simple and cost-effective but rather difficult to optimize RAPD fingerprinting could be used to evaluate genetic and epidemiological relationships of CA-MRSA isolates on condition that the patterns are obtained from carefully optimized laboratory tests.

No MeSH data available.