Limits...
Molecular Investigation of Quinolone Resistance of Quinolone Resistance-Determining Region in Streptococcus pneumoniae Strains Isolated from Iran Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Method.

Kargar M, Moein Jahromi F, Doosti A, Handali S - Osong Public Health Res Perspect (2014)

Bottom Line: The resistance of Streptococcus pneumoniae to the recently available antibiotic treatment has been a growing problem.Antibiotic susceptibility test was performed using the CLSI (Clinical and Laboratory Standards Institute) criteria on three different generations of quinolone family, with nalidixic acid (82.22%) showing the highest resistance and levofloxacin (42.22%) the least resistance.Results indicated that there is a significant correlation between quinolone resistance development and mutations in the parE gene as well as in the parC and gyrA genes.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Islamic Azad University, Jahrom Branch, Jahrom, Iran.

ABSTRACT

Objectives: The resistance of Streptococcus pneumoniae to the recently available antibiotic treatment has been a growing problem. The aim of the study was to determine the quinolone-resistant strains and detect the presence of mutations in the quinolone resistance-determining regions of the gyrA, parE, and parC genes.

Methods: In this study, for the first time in Iran, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to investigate the presence of mutations at quinolone resistance-determining regions of topoisomerase IV and DNA gyrase on 82 S. pneumoniae strains, among them 45 clinical samples were from patients and 37 from healthy carriers (control group).

Results: In clinical samples, 34 (75.56%) strains contained mutations in the parC gene, 31 (68.89%) carried mutations in the gyrA gene, and 14 (31.11%) had parE gene mutations. Antibiotic susceptibility test was performed using the CLSI (Clinical and Laboratory Standards Institute) criteria on three different generations of quinolone family, with nalidixic acid (82.22%) showing the highest resistance and levofloxacin (42.22%) the least resistance.

Conclusion: Results indicated that there is a significant correlation between quinolone resistance development and mutations in the parE gene as well as in the parC and gyrA genes.

No MeSH data available.


Related in: MedlinePlus

PCR-RFLP, parE gene. (A) M, represents 100 bp DNA marker; 1. negative control; and 2, uncut PCR product (290 bp). (B) M, represents 100 bp DNA marker; 1, MspI restriction enzyme digestion products of wild-type strain giving 160 bp and 130 bp fragments; and 2, products of mutant strain digestion by MspI enzyme representing 130 bp, 130 bp, and 30 bp fragments. PCR = polymerase chain reaction; RFLP = restriction fragment length polymorphism.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4225646&req=5

fig2: PCR-RFLP, parE gene. (A) M, represents 100 bp DNA marker; 1. negative control; and 2, uncut PCR product (290 bp). (B) M, represents 100 bp DNA marker; 1, MspI restriction enzyme digestion products of wild-type strain giving 160 bp and 130 bp fragments; and 2, products of mutant strain digestion by MspI enzyme representing 130 bp, 130 bp, and 30 bp fragments. PCR = polymerase chain reaction; RFLP = restriction fragment length polymorphism.

Mentions: The results indicated that 34 (75.55%) out of 45 clinical isolates contained mutations in the parC gene, 31 (68.89%) had mutations in the gyrA gene, and 14 (31.11%) had mutations in the parE gene. Moreover, from 37 control samples, nine (24.32%) strains with mutations in the parC gene, seven (18.99%) with mutations in the gyrA gene, and eight (21.52%) with mutations in the parE gene were observed. Detection of three DNA fragments of 200 bp, 80 bp, and 80 bp, as a result of digestion of the 360 bp parC gene amplicons with Sau3A enzyme, represented the existence of mutations, whereas the observation of two fragments of 200 bp and 160 bp indicated the absence of mutations in the correspondent gene (Figure 1). Furthermore, the mutant parE gene gave three fragments of 130 bp, 130 bp, and 30 bp after digestion of the 290 bp amplicons with MspI enzyme, where detection of two fragments of 130 bp and 160 bp suggested the absence of mutations in this gene (Figure 2). Detection of three DNA fragments of 180 bp, 140 bp, and 60 bp as a result of digestion of the 380 bp gyrA gene amplicons with AluI enzyme represented the existence of mutations, whereas the observation of two fragments of 200 bp and 180 bp indicated the absence of mutations in the correspondent gene (Figure 3).


Molecular Investigation of Quinolone Resistance of Quinolone Resistance-Determining Region in Streptococcus pneumoniae Strains Isolated from Iran Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Method.

Kargar M, Moein Jahromi F, Doosti A, Handali S - Osong Public Health Res Perspect (2014)

PCR-RFLP, parE gene. (A) M, represents 100 bp DNA marker; 1. negative control; and 2, uncut PCR product (290 bp). (B) M, represents 100 bp DNA marker; 1, MspI restriction enzyme digestion products of wild-type strain giving 160 bp and 130 bp fragments; and 2, products of mutant strain digestion by MspI enzyme representing 130 bp, 130 bp, and 30 bp fragments. PCR = polymerase chain reaction; RFLP = restriction fragment length polymorphism.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4225646&req=5

fig2: PCR-RFLP, parE gene. (A) M, represents 100 bp DNA marker; 1. negative control; and 2, uncut PCR product (290 bp). (B) M, represents 100 bp DNA marker; 1, MspI restriction enzyme digestion products of wild-type strain giving 160 bp and 130 bp fragments; and 2, products of mutant strain digestion by MspI enzyme representing 130 bp, 130 bp, and 30 bp fragments. PCR = polymerase chain reaction; RFLP = restriction fragment length polymorphism.
Mentions: The results indicated that 34 (75.55%) out of 45 clinical isolates contained mutations in the parC gene, 31 (68.89%) had mutations in the gyrA gene, and 14 (31.11%) had mutations in the parE gene. Moreover, from 37 control samples, nine (24.32%) strains with mutations in the parC gene, seven (18.99%) with mutations in the gyrA gene, and eight (21.52%) with mutations in the parE gene were observed. Detection of three DNA fragments of 200 bp, 80 bp, and 80 bp, as a result of digestion of the 360 bp parC gene amplicons with Sau3A enzyme, represented the existence of mutations, whereas the observation of two fragments of 200 bp and 160 bp indicated the absence of mutations in the correspondent gene (Figure 1). Furthermore, the mutant parE gene gave three fragments of 130 bp, 130 bp, and 30 bp after digestion of the 290 bp amplicons with MspI enzyme, where detection of two fragments of 130 bp and 160 bp suggested the absence of mutations in this gene (Figure 2). Detection of three DNA fragments of 180 bp, 140 bp, and 60 bp as a result of digestion of the 380 bp gyrA gene amplicons with AluI enzyme represented the existence of mutations, whereas the observation of two fragments of 200 bp and 180 bp indicated the absence of mutations in the correspondent gene (Figure 3).

Bottom Line: The resistance of Streptococcus pneumoniae to the recently available antibiotic treatment has been a growing problem.Antibiotic susceptibility test was performed using the CLSI (Clinical and Laboratory Standards Institute) criteria on three different generations of quinolone family, with nalidixic acid (82.22%) showing the highest resistance and levofloxacin (42.22%) the least resistance.Results indicated that there is a significant correlation between quinolone resistance development and mutations in the parE gene as well as in the parC and gyrA genes.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Islamic Azad University, Jahrom Branch, Jahrom, Iran.

ABSTRACT

Objectives: The resistance of Streptococcus pneumoniae to the recently available antibiotic treatment has been a growing problem. The aim of the study was to determine the quinolone-resistant strains and detect the presence of mutations in the quinolone resistance-determining regions of the gyrA, parE, and parC genes.

Methods: In this study, for the first time in Iran, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to investigate the presence of mutations at quinolone resistance-determining regions of topoisomerase IV and DNA gyrase on 82 S. pneumoniae strains, among them 45 clinical samples were from patients and 37 from healthy carriers (control group).

Results: In clinical samples, 34 (75.56%) strains contained mutations in the parC gene, 31 (68.89%) carried mutations in the gyrA gene, and 14 (31.11%) had parE gene mutations. Antibiotic susceptibility test was performed using the CLSI (Clinical and Laboratory Standards Institute) criteria on three different generations of quinolone family, with nalidixic acid (82.22%) showing the highest resistance and levofloxacin (42.22%) the least resistance.

Conclusion: Results indicated that there is a significant correlation between quinolone resistance development and mutations in the parE gene as well as in the parC and gyrA genes.

No MeSH data available.


Related in: MedlinePlus