scholarly journals Amplified fragment length polymorphism analysis of Mycobacterium avium complex isolates recovered from southern California

2007 ◽  
Vol 56 (9) ◽  
pp. 1152-1160 ◽  
Author(s):  
Stacy L. Pfaller ◽  
Timothy W. Aronson ◽  
Alan E. Holtzman ◽  
Terry C. Covert
Keyword(s):  
Mycobacterium Avium ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Southern California ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Amplified Fragment Length Polymorphism ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Environmental Isolates

Fine-scale genotyping methods are necessary in order to identify possible sources of human exposure to opportunistic pathogens belonging to the Mycobacterium avium complex (MAC). In this study, amplified fragment length polymorphism (AFLP) analysis was evaluated for fingerprinting 159 patient and environmental MAC isolates from southern California. AFLP analysis accurately identified strains belonging to M. avium and Mycobacterium intracellulare and differentiated between strains within each species. The method was also able to differentiate strains that were presumed to be genetically identical in two previous studies using large RFLP analysis with PFGE, or PCR-amplification of DNA segments located between insertion sequences IS1245 and IS1311. For M. avium, drinking-water isolates clustered more closely with each other than with patient or food isolates. Patient isolates were more genetically diverse. None of the environmental isolates shared identical AFLP patterns with patient isolates for either species. There were, however, environmental isolates that shared identical patterns, and patient isolates that shared identical patterns. A subset of the isolates, which are referred to as MX isolates due to their ambiguous identification with the Gen-Probe system, produced AFLP patterns similar to those obtained from M. intracellulare isolates. Sequence analysis of 16S rDNA obtained from the MX isolates suggests that they are strains of M. intracellulare that were not correctly identified by the M. intracellulare AccuProbe from Gen-Probe.

scholarly journals Genomic Relatedness of ChlamydiaIsolates Determined by Amplified Fragment Length Polymorphism Analysis

1999 ◽  
Vol 181 (15) ◽  
pp. 4469-4475 ◽  
Author(s):  
Adam Meijer ◽  
Servaas A. Morré ◽  
Adriaan J. C. Van Den Brule ◽  
Paul H. M. Savelkoul ◽  
Jacobus M. Ossewaarde
Keyword(s):  
Cluster Analysis ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Amplified Fragment Length Polymorphism ◽  
Chlamydia Psittaci ◽  
Rrna Genes ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Genomic Relatedness

ABSTRACT The genomic relatedness of 19 Chlamydia pneumoniaeisolates (17 from respiratory origin and 2 from atherosclerotic origin), 21 Chlamydia trachomatis isolates (all serovars from the human biovar, an isolate from the mouse biovar, and a porcine isolate), 6 Chlamydia psittaci isolates (5 avian isolates and 1 feline isolate), and 1 Chlamydia pecorum isolate was studied by analyzing genomic amplified fragment length polymorphism (AFLP) fingerprints. The AFLP procedure was adapted from a previously developed method for characterization of clinical C. trachomatis isolates. The fingerprints of all C. pneumoniae isolates were nearly identical, clustering together at a Dice similarity of 92.6% (± 1.6% standard deviation). The fingerprints of the C. trachomatis isolates of human, mouse, and swine origin were clearly distinct from each other. The fingerprints of the isolates from the human biovar could be divided into at least 12 different types when the presence or absence of specific bands was taken into account. The C. psittacifingerprints could be divided into a parakeet, a pigeon, and a feline type. The fingerprint of C. pecorum was clearly distinct from all others. Cluster analysis of selected isolates from all species revealed groups other than those based on sequence data from single genes (in particular, omp1 and rRNA genes) but was in agreement with available DNA-DNA hybridization data. In conclusion, cluster analysis of AFLP fingerprints of representatives of all species provided suggestions for a grouping of chlamydiae based on the analysis of the whole genome. Furthermore, genomic AFLP analysis showed that the genome of C. pneumoniae is highly conserved and that no differences exist between isolates of respiratory and atherosclerotic origins.

Comparative Fingerprinting Analysis ofCampylobacter jejuni subsp. jejuni Strains by Amplified-Fragment Length Polymorphism Genotyping

2000 ◽  
Vol 38 (9) ◽  
pp. 3379-3387 ◽  
Author(s):  
Bjørn-Arne Lindstedt ◽  
Even Heir ◽  
Traute Vardund ◽  
Kjetil K. Melby ◽  
Georg Kapperud
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Amplified Fragment Length Polymorphism ◽  
Epidemiological Surveillance ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Ofcampylobacter Jejuni

Amplified-fragment length polymorphism (AFLP) analysis with the endonucleases BglII and MfeI was used to genotype 91 Campylobacter jejuni subsp. jejunistrains from outbreaks and sporadic cases. AFLP-generated fragments were labeled with fluorescent dye and separated by capillary electrophoresis. The software packages GeneScan and GelCompar II were used to calculate AFLP pattern similarities and to investigate phylogenetic relationships among the genotyped strains. The AFLP method was compared with two additional DNA-based typing methods, pulsed-field gel electrophoresis (PFGE) using SmaI and restriction fragment length polymorphism analysis on PCR products (PCR-RFLP) of theflaA and flaB genes. We found that AFLP analysis of C. jejuni strains is a rapid method that offers better discriminatory power than do both PFGE and PCR-RFLP. AFLP and, to a lesser extent, PCR-RFLP could differentiate strains within the same PFGE profiles, which also makes PCR-RFLP an alternative to PFGE. We were able to clearly distinguish 9 of 10 recognized outbreaks by AFLP and to identify similarities among outbreak and sporadic strains. Therefore, AFLP is suitable for epidemiological surveillance ofC. jejuni and will be an excellent tool for source identification in outbreak situations.

scholarly journals Amplified Fragment Length Polymorphism Analysis Provides Strategies for Improvement of Bitter Gourd (Momordica charantia L.)

HortScience ◽  
2008 ◽  
Vol 43 (1) ◽  
pp. 127-133 ◽  
Author(s):  
Ambika B. Gaikwad ◽  
Tusar Kanti Behera ◽  
Anand K. Singh ◽  
Devanshi Chandel ◽  
Jawahir L. Karihaloo ◽  
...  
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Genetic Relationships ◽  
Amplified Fragment Length Polymorphism ◽  
Momordica Charantia ◽  
Bitter Gourd ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Diversity Assessment

Monoecious bitter gourd (Momordica charantia L. var. minima and maxima Williams & Ng), a cucurbit of major economic importance, is widely cultivated in India, China, Africa, and South America. Although the morphology (i.e., growth habit and fruit shape, size, color, and surface texture) of Indian bitter gourd is diverse and gynoecious sex forms exist, a comprehensive diversity assessment of ecotypes has not been performed. Therefore, the genetic relatedness of 38 Indian cultigens (commercial varieties and cultivated landraces originating from different agroecological zones) was determined by amplified fragment length polymorphism (AFLP) analysis. Six primer combinations yielded a total of 519 bands of which 404 (77.8%) were polymorphic among the cultigens examined. Unweighted pair group cluster analyses were performed using Jaccard's genetic similarities to define genetic relationships among cultigens. Genetic similarities among cultigens ranged between 0.44 and 0.88, indicating that the bitter gourd cultigens examined were genetically diverse. Moreover, putative AFLP loci defined genetic relationships that allowed for partitioning of cultigens into two distinct groups [Group 1 and Group II (node 1); bootstrap = 100%] after cluster analysis. With rare exception, cultigens were grouped with respect to geographical region, in which cultigens within a group and subgroups possessed high degrees of genetic similarity. The relatively high marker indices (6.2 to 19.4), polymorphic information content of the markers used (0.20 to 0.25), and multiplex ratios (28.9 to 77.4) collectively indicate that the AFLP markers used are discriminatory in bitter gourd and that the analysis of the broad-based cultigens described provides valuable baseline information for advancing initial breeding strategies for this crop species.

Amplified fragment length polymorphism analysis of different geographic populations of the gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae)

1999 ◽  
Vol 89 (1) ◽  
pp. 79-88 ◽  
Author(s):  
A. Reineke ◽  
P. Karlovsky ◽  
C.P.W. Zebitz
Keyword(s):  
Cluster Analysis ◽  
Length Polymorphism ◽  
Gypsy Moth ◽  
Lymantria Dispar ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Insect Pests ◽  
Amplified Fragment Length Polymorphism ◽  
Aflp Analysis ◽  
Polymorphism Analysis

AbstractThe gypsy moth, Lymantria dispar Linnaeus, is one of the most serious insect pests of palaearctic and nearctic hardwood forests. We used amplified fragment length polymorphism (AFLP) to detect genetic diversity within and among gypsy moth populations. Five AFLP primer combinations were used on 98 L. dispar samples from different parts of Europe, Asia and North America, detecting a total of 481 polymorphic and 58 monomorphic fragments. Genetic similarities based on these data were calculated and cluster analysis was performed to graphically display groupings between isolates. Lymantria dispar individuals from close geographical areas of Europe were mostly grouped together in cluster analysis resulting in the formation of subgroups corresponding to the origin of the samples. Supporting this observation, clustering of individuals from 22 neighbouring populations in southern Germany agreed well with the region they originated from. Thus, AFLP analysis revealed the existence of a certain degree of genetic variability between European gypsy moth populations that could be explained by the accumulation of polymorphisms resulting from both historical population bottlenecks and the adaptation to different environmental conditions. The results of this study therefore demonstrate that AFLP analysis is a sensitive technique for distinguishing genotypes from different geographic origins as well as from neighbouring local populations and provides sufficient molecular markers for future characterization of the gypsy moth genome.

scholarly journals Investigating Parentage and Hybridity of Three Azaleodendrons Using Amplified Fragment Length Polymorphism Analysis

HortScience ◽  
2007 ◽  
Vol 42 (3) ◽  
pp. 740-743 ◽  
Author(s):  
Ryan N. Contreras ◽  
Thomas G. Ranney ◽  
Susana R. Milla-Lewis ◽  
G. Craig Yencho
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Genetic Similarity ◽  
Morphological Analysis ◽  
Fragment Length Polymorphism ◽  
Amplified Fragment Length Polymorphism ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Single Clone ◽  
Similar Appearance

Morphological analysis historically has been used to determine parentage of unknown hybrids. This can be difficult when potential parents have similar appearance, as in the case of three azaleodendron cultivars, Rhododendron L. ‘Fragrans’, ‘Fragrans Affinity’, and ‘Fragrant Affinity’. These cultivars are similar in name and appearance, and all are purported hybrids of R. catawbiense Michx. or R. ponticum L. and R. viscosum (L.) Torr. Amplified fragment length polymorphism (AFLP) analysis was conducted to determine whether the cultivars are synonyms or distinct clones and to elucidate the parental species. The three cultivars, suspected to be hybrids between taxa in subgenera Hymenanthes (Blume) K.Koch (evergreen rhododendrons) and Pentanthera (G.Don) Pojarkova (deciduous azaleas), and related taxa from each subgenus were evaluated using 31 AFLP primer combinations. Genetic similarity, calculated using Jaccard's coefficient, among the hybrids ranged from 53% to 71%, indicating that they are distinct cultivars and not a single clone. Genetic similarity was highest between the hybrids and R. ponticum among the evergreen rhododendrons, and R. viscosum among the deciduous azaleas. A dendrogram generated using the genetic similarity matrix grouped taxa into their respective subgenera, with the three cultivars nested intermediately between subgenera but more closely with subgenus Hymenanthes and particularly R. ponticum, suggesting it is the evergreen rhododendron parent. Furthermore, principle components grouped R. ponticum more closely with the hybrids and there were 18 AFLP fragments unique to R. ponticum and the hybrids. However, no unique AFLP bands were shared exclusively among the hybrids and the purported deciduous azalea parent, R. viscosum, suggesting that the original azalea parents may have been hybrids.

scholarly journals Efficient DNA Fingerprinting of Clostridium botulinum Types A, B, E, and F by Amplified Fragment Length Polymorphism Analysis

2005 ◽  
Vol 71 (3) ◽  
pp. 1148-1154 ◽  
Author(s):  
Riikka Keto-Timonen ◽  
Mari Nevas ◽  
Hannu Korkeala
Keyword(s):  
Dna Fingerprinting ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Clostridium Botulinum ◽  
Fragment Length Polymorphism ◽  
Group Identification ◽  
Amplified Fragment Length Polymorphism ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Group I

ABSTRACT Amplified fragment length polymorphism (AFLP) analysis was applied to characterize 33 group I and 37 group II Clostridium botulinum strains. Four restriction enzyme and 30 primer combinations were screened to tailor the AFLP technique for optimal characterization of C. botulinum. The enzyme combination HindIII and HpyCH4IV, with primers having one selective nucleotide apiece (Hind-C and Hpy-A), was selected. AFLP clearly differentiated between C. botulinum groups I and II; group-specific clusters showed <10% similarity between proteolytic and nonproteolytic C. botulinum strains. In addition, group-specific fragments were detected in both groups. All strains studied were typeable by AFLP, and a total of 42 AFLP types were identified. Extensive diversity was observed among strains of C. botulinum type E, whereas group I had lower genetic biodiversity. These results indicate that AFLP is a fast, highly discriminating, and reproducible DNA fingerprinting method with excellent typeability, which, in addition to its suitability for typing at strain level, can be used for C. botulinum group identification.

scholarly journals Fluorescent Amplified Fragment Length Polymorphism Analysis of Norwegian Bacillus cereus and Bacillus thuringiensis Soil Isolates

2001 ◽  
Vol 67 (10) ◽  
pp. 4863-4873 ◽  
Author(s):  
Lawrence O. Ticknor ◽  
Anne-Brit Kolstø ◽  
Karen K. Hill ◽  
Paul Keim ◽  
Miriam T. Laker ◽  
...  
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Amplified Fragment Length Polymorphism ◽  
Aflp Analysis ◽  
Rrna Gene ◽  
Polymorphism Analysis ◽  
Entire Study ◽  
Genetic Groups ◽  
Reference Strains

ABSTRACT We examined 154 Norwegian B. cereus andB. thuringiensis soil isolates (collected from five different locations), 8 B. cereus and 2B. thuringiensis reference strains, and 2Bacillus anthracis strains by using fluorescent amplified fragment length polymorphism (AFLP). We employed a novel fragment identification approach based on a hierarchical agglomerative clustering routine that identifies fragments in an automated fashion. No method is free of error, and we identified the major sources so that experiments can be designed to minimize its effect. Phylogenetic analysis of the fluorescent AFLP results reveals five genetic groups in these group 1 bacilli. The ATCC reference strains were restricted to two of the genetic groups, clearly not representative of the diversity in these bacteria. Both B. anthracis strains analyzed were closely related and affiliated with a B. cereus milk isolate (ATCC 4342) and a B. cereus human pathogenic strain (periodontitis). Across the entire study, pathogenic strains, including B. anthracis, were more closely related to one another than to the environmental isolates. Eight strains representing the five distinct phylogenetic clusters were further analyzed by comparison of their 16S rRNA gene sequences to confirm the phylogenetic status of these groups. This analysis was consistent with the AFLP analysis, although of much lower resolution. The innovation of automated genotype analysis by using a replicated and statistical approach to fragment identification will allow very large sample analyses in the future.

scholarly journals Amplified Fragment Length Polymorphism Analysis ofCampylobacter jejuni Strains Isolated from Chickens and from Patients with Gastroenteritis or Guillain-Barr� or Miller Fisher Syndrome

2000 ◽  
Vol 66 (9) ◽  
pp. 3917-3923 ◽  
Author(s):  
Birgitta Duim ◽  
C. Wim Ang ◽  
Alex van Belkum ◽  
Alan Rigter ◽  
Nan W. J. van Leeuwen ◽  
...  
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Genetic Relationships ◽  
Amplified Fragment Length Polymorphism ◽  
Miller Fisher Syndrome ◽  
Group Method ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Fisher Syndrome

ABSTRACT The high-resolution genotyping method of amplified fragment length polymorphism (AFLP) analysis was used to study the genetic relationships between Campylobacter jejuni strains infecting chickens (n = 54) and those causing gastroenteritis in humans (n = 53). In addition,C. jejuni strains associated with the development of Guillain-Barr� syndrome (GBS) (n = 14) and Miller Fisher syndrome (MFS) (n = 4), two related acute paralytic syndromes in human, were included. Strains were isolated between 1989 and 1998 in The Netherlands. The AFLP banding patterns were analyzed with correlation-based and band-based similarity coefficients and UPGMA (unweighted pair group method using average linkages) cluster analysis. All C. jejuni strains showed highly heterogeneous fingerprints, and no fingerprints exclusive for chicken strains or for human strains were obtained. All strains were separated in two distinct genetic groups. In group A the percentage of human strains was significantly higher and may be an indication that genotypes of this group are more frequently associated with human diseases. We conclude that C. jejuni from chickens cannot be distinguished from human strains and that GBS or MFS related strains do not belong to a distinct genetic group.

scholarly journals Amplified Fragment Length Polymorphism to Detect Clonal Diversity and Distribution of Mycobacterium Avium Subspecies Paratuberculosis in Selected Minnesota Dairy Cattle

2005 ◽  
Vol 17 (4) ◽  
pp. 311-315 ◽  
Author(s):  
Laura A. Kiehnbaum ◽  
Alongkorn Amonsin ◽  
Scott J. Wells ◽  
Vivek Kapur
Keyword(s):  
Mycobacterium Avium ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Molecular Ecology ◽  
Clonal Diversity ◽  
Amplified Fragment Length Polymorphism ◽  
The United States ◽  
Aflp Analysis

The molecular ecology of Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne's disease, is not well understood in the United States. In this study, a DNA fingerprinting method, amplified fragment length polymorphism (AFLP), was used to subtype the pathogen and assess the clonal diversity of MAP in Minnesota dairy herds. Fifty-six fecal culture test–positive isolates from various Minnesota counties and culture dates were analyzed in this study. The AFLP identified 11 profiles with 50% of isolates representing 1 major profile. The major profile was distributed across the state. The genetic diversity of bovine MAP clones in Minnesota based on AFLP analysis of this data appears to be relatively low.

scholarly journals Evaluation of Amplified Fragment Length Polymorphism Analysis for Inter- and Intraspecific Differentiation ofMycobacterium bovis, M. tuberculosis, andM. ulcerans

2000 ◽  
Vol 38 (10) ◽  
pp. 3675-3680 ◽  
Author(s):  
G. Huys ◽  
L. Rigouts ◽  
K. Chemlal ◽  
F. Portaels ◽  
J. Swings
Keyword(s):  
Correlation Coefficient ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Aflp Marker ◽  
Genotypic Variation ◽  
Amplified Fragment Length Polymorphism ◽  
Resolving Power ◽  
Aflp Analysis ◽  
Polymorphism Analysis

The usefulness of amplified fragment length polymorphism (AFLP) analysis was evaluated for the discrimination of Mycobacterium bovis (17 strains), M. tuberculosis (15 strains), andM. ulcerans (12 strains) at the inter- and intraspecific level. The AFLP technique is a whole-genome coverage genotypic fingerprinting method based on the selective PCR amplification of modified restriction fragments obtained through a double enzymatic digest and subsequent ligation of double-stranded restriction site-specific adapter oligonucleotides. Selective amplification ofApaI/TaqI templates with primer combination A02-T02 (both having an additional C at their 3′ end) generated autoradiographic AFLP fingerprints that were grouped by numerical analysis in two main AFLP clusters allowing clear separation ofM. ulcerans (cluster I) from the M. tuberculosis complex members M. bovis and M. tuberculosis (cluster II). Calculation of similarities using the band-based Dice correlation coefficient instead of the Pearson product-moment correlation coefficient revealed a further subgrouping in cluster II. The two resulting subclusters corresponded with the phenotypic identity of M. bovis and M. tuberculosis, respectively, and could also be visually identified by two AFLP marker bands. Because of the relatively low degree of genotypic variation among the AFLP band patterns of the latter two taxa, no correlation could be found with previously reported molecular typing data or with geographical origin. The use of primer combination A02-T01 (the latter having an A as selective base) did not increase the resolving power within the M. tuberculosis complex but resulted in a visual subgrouping of the M. ulcerans strains that was not observed with primer combination A02-T02. Based on the presence or absence of a single AFLP marker band, the M. ulcerans isolates could be unambiguously classified in two continental types corresponding with the African and Australian origin of the strains, respectively. In conclusion, the radioactive AFLP method proved to be a reproducible and reliable taxonomic tool for the differentiation of the three mycobacterial species under study and also demonstrated its potential use for typing of M. ulceransstrains when employing multiple primer combinations.

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