Application of polymerase chain reaction to fingerprinting Aspergillus fumigatus by random amplification of polymorphic DNA

Loudon, K W; Burnie, J P; Coke, A P; Matthews, Ruth

doi:10.1128/jcm.31.5.1117-1121.1993

Cited by 98 publications

(51 citation statements)

References 22 publications

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“…Both RAPD and RFLP analysis were repeated (data not shown), but although minor variations in band intensity were observed, the overall banding pattern and interpretation were consistent. Previous comparisons between R108-primed RAPD and RFLP typing methods have shown both full agreement (19) and discordance (1,11). Factors such as buffer composition, primer concentration, thermal cycler performance, and gel electrophoresis time may play an important role in the band pattern and affect the discriminatory power of the primer.…”

mentioning

confidence: 98%

Genotypic characterization of sequential Aspergillus fumigatus isolates from patients with cystic fibrosis

Verweij¹,

Meis²,

Sarfati³

et al. 1996

J Clin Microbiol

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Twenty-three sequential Aspergillus fumigatus sputum isolates, which had been collected over a period of 2 years, from two patients with cystic fibrosis were genotyped by random amplified polymorphic DNA PCR and restriction fragment length polymorphism analysis. In patient B, one genotype was predominantly present in the sputum samples, while in the other patient up to nine different genotypes were identified. This study suggests that different patterns of colonization with A. fumigatus exist in patients with cystic fibrosis.

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mentioning

confidence: 98%

Genotypic characterization of sequential Aspergillus fumigatus isolates from patients with cystic fibrosis

Verweij¹,

Meis²,

Sarfati³

et al. 1996

J Clin Microbiol

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“…Similarly, using XbaI, Burnie et al (5) detected six REA types among 21 isolates. Two independent investigations, in which different sets of oligonucleotide primers were used, demonstrated that the random amplified polymorphic DNA (RAPD) method generated a relatively high degree of discrimination among isolates (1,21). Regrettably, these typing methods are not ideal.…”

mentioning

confidence: 99%

“…Few investigators have directly compared the results for typing isolates of A. fumigatus by several different methods. Such studies have used small numbers of isolates, and the division of strains into groups may have been influenced by phenotypic variability (5,21). In contrast, our investigation directly compares the epidemiologic utilities of three genomic typing methods, REA, IEA, and RAPD analysis, applying them to a large collection of A. fumigatus clinical and environmental isolates.…”

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confidence: 99%

Comparison of three typing methods for clinical and environmental isolates of Aspergillus fumigatus

et al. 1995

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To evaluate procedures used for epidemiologic analysis of outbreaks of aspergillosis, we analyzed a collection of 35 Aspergillus fumigatus isolates using three typing methods: isoenzyme analysis (IEA), random amplified polymorphic DNA (RAPD) analysis, and restriction endonuclease analysis (REA). Twenty-one isolates were from a single hospital, with four isolates coming from different patients. Three clinical isolates came from a different hospital, and 11 clinical or environmental isolates were derived from a culture collection. With IEA, the patterns of alkaline phosphatase, esterase, and catalase discriminated nine types. In contrast, 22 types were obtained with five different RAPD primers, and 21 types could be detected with three of these (R108, R151, and UBC90). Restriction endonuclease analysis of genomic DNA, digested with either XbaI, XhoI, or SalI, detected 3, 17, and 13 different REA types, respectively, and 22 types were identified by combining the data from the XhoI and SalI REAs. Twenty-eight types were obtainable with a combination of REA, IEA, and RAPD patterns. Overall, the results pointed to substantial genetic variation among the isolates. Though two isolates had markedly distinct genotypes, their morphologic features and exoantigens were consistent with their being A. fumigatus. The analysis will help in planning epidemiologic studies of aspergillosis.

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“…[19][20][21][22][23][24][25] Several other primers suitable for typing Aspergillus species have later been described by other groups as well. [26][27][28][29] Restriction fragment length polymorphism without hybridrisation When genomic DNA from an Aspergillus isolate is cut with a restriction enzyme with a 6 bp recognition sequence, up to 10 000 different DNA fragments may be obtained. These DNA fragments are on average some 4000 bp long.…”

Section: Random Amplified Polymorphic Dnamentioning

confidence: 99%

Molecular typing of Aspergillus species

Valk

Klaassen

Meis

2008

Mycoses

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Aspergillus species are widely distributed fungi that release large amounts of airborne conidia, which are dispersed in the environment. Several Aspergillus species have been described as human pathogens. Molecular techniques have been developed to investigate the epidemiological relation between environmental and clinical isolates. Several typing methods have been described for Aspergillus species, most of them with reference to Aspergillus fumigatus. Here, we summarise all the different available molecular typing techniques for Aspergillus. The performance of these techniques is evaluated with respect to their practical feasibility, and their interpretation and discriminatory power assessed. For A. fumigatus isolates, a large extent of genetic variability is demonstrated and therefore fingerprinting techniques with high discriminatory power and high reproducibility are required for this species. Afut1-restriction fragment length polymorphism and microsatellite typing showed the highest discriminatory power. In addition, the microsatellites show excellent reproducibility. Other typing techniques are still useful for smaller epidemiological problems and for less well-equipped laboratories.

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Application of polymerase chain reaction to fingerprinting Aspergillus fumigatus by random amplification of polymorphic DNA

Cited by 98 publications

References 22 publications

Genotypic characterization of sequential Aspergillus fumigatus isolates from patients with cystic fibrosis

Genotypic characterization of sequential Aspergillus fumigatus isolates from patients with cystic fibrosis

Comparison of three typing methods for clinical and environmental isolates of Aspergillus fumigatus

Molecular typing of Aspergillus species

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