Typing of Actinomycetes by DNA amplification techniques & Reliability of AP-PCR and ERIC-PCR for typing Acinetobacter baumannii isoaltes from different sources (original) (raw)
Related papers
Infection and Drug Resistance, 2020
Background: Actinomycetes widely exist in nature and these species cause infections in immunocompromised and healthy patients, although they are frequently found as members of the normal microbiota of humans and animals. These subsequent infections are often misdiagnosed as malignancy and tuberculosis. Due to this issue, the present study aimed to determine the presence and diversity of actinomycetes species causing infections in Iranian patients. Materials and Methods: A total of 79 clinical samples collected from five hospitals in Markazi province were analyzed for the existence of actinomycetes using standard protocols for isolation and characterization of the isolates. The conventional tests were used for preliminary identification, the PCR amplification of hsp65 gene, the specific region of the 16S rRNA, and sequence analyses of 16S rRNA were applied for the genus and species identification. MICs of the antimicrobial agent were determined by the broth microdilution method and interpreted according to the NCCLS guidelines. Results: A total of 17 (21.51%) actinomycetes isolates were recovered from clinical samples. In other analyzed samples, eight (10.12%) gram-positive, 12 (15.18) gramnegative bacteria, and six (7.6) fungi isolates were recovered. The most prevalent actinomycetes species were M. fortuitum (17.64%), N. Mexicana and S. heliomycini (11.76% each), and 10 species, ie, N. farcinica, M. lehmannii, M. flavescens, Arthrobacter crystalopoetis, N. neocaledoniensis, M. phocaicum, M. abscessus, M. arupense, M. setense, and N. cyriacigeorgica made up the single isolates. Results of DST illustrated that all of the isolates were susceptible to Amikacin, Levofloxacin, Ofloxacin, and Ciprofloxacin, whereas all of them were resistant to Rifampicin and Doxycycline. Conclusion: In conclusion, increasing isolation of actinomycetes found in various clinical cases merits special attention by health authorities in developing countries. In health centers, action should be taken to increase awareness of appropriate diagnostic criteria and management guidelines for actinomycetes diseases. Furthermore, an increase in the number as well as the quality of national and regional reference laboratories may facilitate more accurate diagnosis of actinomycetes diseases.
Characterization of serologically nontypeable Actinobacillus actinomycetemcomitans isolates
Journal of clinical microbiology, 1998
Our previous studies have shown that Actinobacillus actinomycetemcomitans isolates of a given arbitrarily primed PCR (AP-PCR) genotype belong to the same serotype (of serotypes a through e). In the present study we investigated whether the AP-PCR genotypes of nonserotypeable A. actinomycetemcomitans isolates match those of the serotypeable isolates. The isolates were additionally characterized by restriction analysis of the apaH PCR amplification products. The material included 75 nonserotypeable and 18 serotypeable A. actinomycetemcomitans isolates from 34 epidemiologically unrelated subjects. The serotypeable isolates were obtained from subjects who also harbored nonserotypeable isolates. Eight AP-PCR genotypes were distinguished among the isolates; six genotypes matched those detected in our previous studies, whereas two genotypes were new. Intraindividually, the A. actinomycetemcomitans isolates produced identical AP-PCR banding patterns, regardless of whether they were serotype...
Infection and Drug Resistance
Background: Actinomycetes widely exist in nature and these species cause infections in immunocompromised and healthy patients, although they are frequently found as members of the normal microbiota of humans and animals. These subsequent infections are often misdiagnosed as malignancy and tuberculosis. Due to this issue, the present study aimed to determine the presence and diversity of actinomycetes species causing infections in Iranian patients. Materials and Methods: A total of 79 clinical samples collected from five hospitals in Markazi province were analyzed for the existence of actinomycetes using standard protocols for isolation and characterization of the isolates. The conventional tests were used for preliminary identification, the PCR amplification of hsp65 gene, the specific region of the 16S rRNA, and sequence analyses of 16S rRNA were applied for the genus and species identification. MICs of the antimicrobial agent were determined by the broth microdilution method and interpreted according to the NCCLS guidelines. Results: A total of 17 (21.51%) actinomycetes isolates were recovered from clinical samples. In other analyzed samples, eight (10.12%) gram-positive, 12 (15.18) gramnegative bacteria, and six (7.6) fungi isolates were recovered. The most prevalent actinomycetes species were M. fortuitum (17.64%), N. Mexicana and S. heliomycini (11.76% each), and 10 species, ie, N. farcinica, M. lehmannii, M. flavescens, Arthrobacter crystalopoetis, N. neocaledoniensis, M. phocaicum, M. abscessus, M. arupense, M. setense, and N. cyriacigeorgica made up the single isolates. Results of DST illustrated that all of the isolates were susceptible to Amikacin, Levofloxacin, Ofloxacin, and Ciprofloxacin, whereas all of them were resistant to Rifampicin and Doxycycline. Conclusion: In conclusion, increasing isolation of actinomycetes found in various clinical cases merits special attention by health authorities in developing countries. In health centers, action should be taken to increase awareness of appropriate diagnostic criteria and management guidelines for actinomycetes diseases. Furthermore, an increase in the number as well as the quality of national and regional reference laboratories may facilitate more accurate diagnosis of actinomycetes diseases.
Correlation of six methods for typing nosocomial isolates of Acinetobacter baumannii
Journal of Medical Microbiology, 1995
A comparative study of biotyping, antimicrobial susceptibility, whole-cell protein analysis, plasmid analysis, pulsed-field gel electrophoresis of chromosomal DNA and polymerase chain reaction with arbitrary primers of Acinetobacter baumannii isolates from three large hospitals was performed to determine the best markers for epidemiological purposes. Ninety-two isolates were included : 38 belonged to a previously described outbreak and 54 were randomly selected from sporadic cases of infection. Biotyping, whole-cell protein and plasmid analysis were the least discriminatory methods, whereas antimicrobial susceptibility and polymerase chain reaction with arbitrary primers showed moderate discriminatory power. Typing based on pulsed-field gel electrophoresis of chromosomal DNA appeared to be the best discriminatory method (discrimination index of 0.9623). The addition of polymerase chain reaction with arbitrary primers or antimicrobial susceptibility to pulsed-field gel electrophoresis of chromosomal DNA did not further increase the discriminatory power.
Comparison of six typing methods for Actinobacillus actinomycetemcomitans
Journal of clinical microbiology, 1994
Actinobacillus actinomycetemcomitans is an important pathogen in the etiology of severe periodontitis. For epidemiological studies on the prevalence of certain pathogenic clones and transmission of this bacterium, adequate typing methods are necessary. The purpose of this study was to compare six different typing methods for A. actinomycetemcomitans. Five reference strains and 27 fresh clinical isolates from periodontitis patients were used. Serotyping showed 12 serotype a strains, 13 type b strains, 6 type c strains, and 1 nontypeable strain. Biotyping on the basis of the fermentation of mannose, mannitol, and xylose resulted in six biotypes. Antibiogram typing was evaluated by measuring the inhibition zones of seven antibiotics in agar diffusion tests. With this method eight main types which could be further differentiated into 15 subtypes were found. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of outer membrane proteins were similar among all isolates teste...
Anaerobe, 2005
In clinical microbiology laboratories, the identification of Actinomyces-like bacteria can be very laborious and problematic. In the present study, we focused on reactivity patterns of 4 commercial test kits, RapID ANA II, RapID 32A, RapID CB Plus, and BBL Crystal ANR ID, that could be used for rapid preliminary identification of Actinomyces isolates belonging to newly described Actinomyces and closely related species. Out of the 54 strains tested, 25 strains (46%) were correctly identified to the genus/group level by BBL Crystal ANR ID system, 16 strains (30%) by RapID 32 A, 11 strains (20%) by RapID CB Plus, and 7 strains (13%) by RapID ANA II. The main problems with these kits were due to occasional weak enzymatic and sugar fermentation reactions. In conclusion, chromogenic substrate sensitivity and specificity need to be enhanced in order to improve the reliability of the test results of these kits, and the present database updated in order to more precisely identify newly described Actinomyces and closely related species.