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At least 973 records · Page 54Linked to original sources
PubMed1994

Transmission of tuberculosis in New York City. An analysis by DNA fingerprinting and conventional epidemiologic methods.

BACKGROUND: The incidence of tuberculosis and drug resistance is increasing in the United States, but it is not clear how much of the increase is due to reactivation of latent infection and how much to recent transmission. METHODS: We performed DNA fingerprinting using restriction-fragment-length polymorphism (RFLP) analysis of at least one isolate from every patient with confirmed tuberculosis at a major hospital in the Bronx, New York, from December 1, 1989, through December 31, 1992. Medical records and census-tract data were reviewed for relevant clinical, social, and demographic data. RESULTS: Of 130 patients with tuberculosis, 104 adults (80 percent) had complete medical records and isolates whose DNA fingerprints could be evaluated. Isolates from 65 patients (62.5 percent) had unique RFLP patterns, whereas isolates from 39 patients (37.5 percent) had RFLP patterns that were identical to those of an isolate from at least 1 other study patient; the isolates in the latter group were classified into 12 clusters. Patients whose isolates were included in one of the clusters were inferred to have recently transmitted disease. Independent risk factors for having a clustered isolate included seropositivity for the human immunodeficiency virus (HIV) (odds ratio for Hispanic patients, 4.31; P = 0.02; for non-Hispanic patients, 3.12; P = 0.07), Hispanic ethnicity combined with HIV seronegativity (odds ratio, 5.13; P = 0.05), infection with drug-resistant tuberculosis (odds ratio, 4.52; P = 0.005), and younger age (odds ratio, 1.59; P = 0.02). Residence in sections of the Bronx with a median household income below $20,000 was also associated with having a clustered isolate (odds ratio, 3.22; P = 0.04). CONCLUSIONS: In the inner-city community we studied, recently transmitted tuberculosis accounts for approximately 40 percent of the incident cases and almost two thirds of drug-resistant cases. Recent transmission of tuberculosis, and not only reactivation of latent disease, contributes substantially to the increase in tuberculosis.

AIDS-Related Opportunistic Infections↗
PubMed1998

The use of DNA fingerprinting to assess monozygotic twinning in Meishan and Landrace x large white pigs.

The extent of embryo mortality is usually estimated by the difference between the numbers of corpora lutea and embryos in the same animal, which assumes that each corpora lutea represents one potential embryo. Recent observations from the authors' laboratory reveal situations in which the number of embryos exceeds the number of corpora lutea, implicating the presence of identical twins. The objective of the present study was to establish DNA fingerprinting techniques to investigate the prevalence of monozygotic twinning in two breeds of pig. DNA fingerprints of every fetus carried by 6 Meishan (MS) and 6 Landrace x Large White (LxLW) gilts on Day 29 +/- 2 of pregnancy were obtained. Five LxLW and 3 MS litters carried no identical fetuses. The remaining gilts carried a pair of fetuses with indistinguishable DNA profiles. No pairs of fetuses were monochorionic. These results suggest that monozygotic twinning in the pig occurs during embryo cleavage or blastocyst development and may be more prevalent in MS compared with indigenous breeds.

Animals↗
PubMed1994

DNA fingerprinting analysis of parent-offspring conflict in a bee.

Demonstrating the importance of haplodiploidy in the evolution of eusociality among the Hymenoptera (bees, wasps, and ants) requires estimation of four parameters: relatedness between cooperating individuals, effective mating frequency, sex ratio, and rates of worker reproduction. Multilocus DNA fingerprinting techniques permitted the precise determination of these parameters for the primitively eusocial bee Augochlorella striata (Halictidae). DNA fingerprints revealed an unprecedented resolution of genetic relationships within colonies, detecting factors such as intraspecific nest parasitism and diploid males that confounded estimates of relatedness and sex ratio, respectively. Parameter estimates (i) corroborate recent evidence for queen-worker conflict over the sex ratio and (ii) implicate the role of haplodiploidy in the evolution of worker behavior.

Animals↗
PubMed1995

Genomic DNA fingerprinting by restriction fragment end labeling.

A typing method for bacteria was developed and applied to several species, including Escherichia coli and Actinobacillus actinomycetemcomitans. Total genomic DNA was digested with a restriction endonuclease, and fragments were enabled with [alpha-32P]dATP by using the Klenow fragment of DNA polymerase and separated by electrophoresis in 6% polyacrylamide/8 M urea (sequencing gel). Depending on the restriction endonuclease and the bacterium, the method produced approximately 30-50 well-separated fragments in the size range of 100-400 nucleotides. For A. actinomycetemcomitans, all strains had bands in common. Nevertheless, many polymorphisms could be observed, and the 31 strains tested could be classified into 29 distinct types. Furthermore, serotype-specific fragments could be assigned for the three serotypes investigated. The method described is very sensitive, allowing more distinct types to be distinguished than other commonly used typing methods. When the method was applied to 10 other clinically relevant bacterial species, both species-specific bands and strain-specific bands were found. Isolates from different locations of one patient showed indistinguishable patterns. Computer-assisted analysis of the DNA fingerprints allowed the determination of similarity coefficients. It is concluded that genomic fingerprinting by restriction fragment end labeling (RFEL) is a powerful and generally applicable technique to type bacterial species.

DNA Fingerprinting↗
PubMed2001

Pyrolysis mass spectrometry (PyMS) and 16S-23S rDNA spacer region fingerprinting suggests the presence of novel acinetobacters in activated sludge.

Screening of large numbers of Acinetobacter spp. from activated sludge systems with Pyrolysis Mass Spectrometry (PyMS) showed that many did not cluster tightly with the currently described genomic species which have been obtained mainly from clinical sources. Selected isolates were then genotypically fingerprinted using their 16S-23S rDNA spacer region, and again the data revealed considerable differences in the genomic fingerprints of many of these activated sludge isolates to the predominantly clinical genomic species. In fact, few could be identified from them. The possibility that the current speciation within this genus is not adequate to encompass all these environmental isolates is addressed in relation to the methods used to study the population dynamics of Acinetobacter in activated sludge.

Acinetobacter↗
PubMed2002

Differentiation of nocardia species by PCR-randomly amplified polymorphic DNA fingerprinting.

Representatives of fifteen validly described and three non-validly described species of Nocardia were assigned to nineteen groups based on an optimised PCR-randomly amplified polymorphic DNA fingerprinting technique. Species specific banding patterns were recognised for the representatives of N. brasiliensis, N. crassostreae, N. farcinica, N. otitidiscaviarum and N. seriola. Unique banding patterns were also seen for the type strains of N. brevicatena, N. carnea, N. salmonicida, N. uniformis and N. vaccinii, and for the single representatives of "N. fusca", "N. pseudosporangifera", and "N. violaceofusca". More than one banding pattern was detected for the N. asteroides, N. flavorosea, N. nova, N. pseudobrasiliensis and N. transvalensis strains though in the case of the representative strains of N. nova and N. transvalensis the patterns were similar for each of these species. The results are in line with current trends in nocardial systematics thereby indicating that PCR-randomly amplified polymorphic DNA fingerprinting provides valuable data for the classification and identification of pathogenic nocardiae to the species level.

DNA Fingerprinting↗
PubMed2004

BOX-pCR fingerprinting as a powerful tool to reveal synonymous names in the genus Streptomyces. Emended descriptions are proposed for the species Streptomyces cinereorectus, S. fradiae, S. tricolor, S. colombiensis, S. filamentosus, S. vinaceus and S. phaeopurpureus.

The type strains of 451 validly described Streptomyces species were screened using the BOX-PCR fingerprint technique. Of the different primers tested, the BOX primer yielded the most robust and reproducible band patterns. The majority of the species (350) had an unique pattern each. A total number of 30 clusters were delineated comprising species with nearly identical patterns. Four of these clusters grouped synonymous Streptomyces species known from previous studies. A high correlation exists between BOX-PCR fingerprinting and DNA-DNA homology data. In the present study, the latter was confirmed for 7 additional BOX clusters and led to the following emended descriptions: S. cochleatus is a subjective synonym of S. cinereorectus, S. roseoflavus of S. fradiae, S. roseodiastaticus of S. tricolor, S. roseosporus of S. filamentosus, S. distallicus of S. colombiensis, S. arabicus of S. vinaceus and S. phaeoviridis of S. phaeopurpureus. Although not yet confirmed by DNA-DNA pairing studies, rep-PCR data suggested the presence of at least 33 additional junior synonyms.

DNA Fingerprinting↗
PubMed2003

Identification of Bifidobacterium species using rep-PCR fingerprinting.

The aim of the present study was to evaluate the use of repetitive DNA element PCR fingerprinting (rep-PCR) for the taxonomic discrimination among the currently described species within the genus Bifidobacterium. After evaluating several primer sets targeting the repetitive DNA elements BOX, ERIC, (GTG)s and REP, the BOXA1R primer was found to be the most optimal choice for the establishment of a taxonomical framework of 80 Bifidobacterium type and reference strains. Subsequently, the BOX-PCR protocol was tested for the identification of 48 unknown bifidobacterial isolates originating from human faecal samples and probiotic products. In conclusion, rep-PCR fingerprinting using the BOXA1R primer can be considered as a promising genotypic tool for the identification of a wide range of bifidobacteria at the species, subspecies and potentially up to the strain level.

Bacterial Typing Techniques↗
PubMed2000

Genotyping Cryptococcus neoformans var. neoformans with specific primers designed from PCR-fingerprinting bands sequenced using a modified PCR-based strategy.

Cryptococcus neoformans var. neoformans can be subdivided into six genotypes (VN1-VN6) based on different combinations of four major bands (420, 475, 540 and 800 bp) obtained by polymerase chain reaction (PCR) fingerprinting using the (GACA)4 primer. The aim of this study was to identify specific primers able to amplify these bands. A modified PCR-based sequencing strategy was adopted to overcome the limitations of using (GACA)4 as a single cycle sequencing primer. The original bands, made up of amplicons with two terminal (GACA)4 sequences, were digested with five restriction enzymes. Digestion products yielding two or three fragments were selected. Each fragment was expected to have no more than one terminal (GACA)4 sequence, making cycle sequencing possible. Fragments were purified and sequenced with the (GACA)4 primer. New primers specific for each of the four major bands were then designed and the remaining regions were sequenced using both purified bands and PCR-fingerprinting products as template. These primers were used to amplify the genomic DNA of 12 C. neoformans strains and five strains of other yeast species. The new primers, used as separate pairs or in a mixture of all pairs, amplified the expected bands only in C. neoformans var. neoformans strains, confirming the species specificity of the bands selected for molecular typing of this yeast.

Base Sequence↗
PubMed1990

Plasmid DNA fingerprinting of Acinetobacter calcoaceticus subspecies anitratus from intubated and mechanically ventilated patients.

Forty-three intubated and mechanically ventilated patients in five intensive care units (ICUs) of one hospital developed respiratory colonization or infection with Acinetobacter calcoaceticus subspecies anitratus over a 16-month interval. Neither the frequency nor rate of A anitratus isolation exceeded the hospital endemic norms. Single isolates from 34 of the patients were subtyped by plasmid DNA analysis, two biotyping systems and antimicrobial susceptibility to 24 drugs. Plasmid DNA fingerprints were distinct in 18 isolates (they differed from each other and all others), similar in two and identical or similar in ten. The latter group of isolates were recovered from patients in four ICUs. Reproducibility of biotyping was poor. Neither biotyping nor antimicrobial susceptibility were successful in identifying sameness among the group isolates nor differences among other isolates. We conclude that plasmid DNA fingerprinting should be used to assess the possibility of multiple patient transmissions of the same A anitratus strain in the absence of an obvious outbreak.

Acinetobacter↗
PubMed2006

DNA fingerprinting in the criminal justice system: an overview.

DNA fingerprinting is a powerful technology that has revolutionized forensic science. No two individuals can have an identical DNA pattern except identical twins. Such DNA-based technologies have enormous social implications and can help in the fight against crime. This technology has experienced many changes over time with many advancements occurring. DNA testing is a matter of serious concern as it involves ethical issues. This article describes various trends in DNA fingerprinting and the current technology used in DNA profiling, possible uses and misuses of DNA databanks and ethical issues involved in DNA testing. Limitations and problems prevailing in this field are highlighted.

Criminal Law↗
PubMed2005

The effects of nuclear DNA content (C-value) on the quality and utility of AFLP fingerprints.

BACKGROUND AND AIMS: Nuclear DNA content (C-value) varies approximately 1000-fold across the angiosperms, and this variation has been reported to have an effect on the quality of AFLP fingerprints. Various methods have been proposed for circumventing the problems associated with small and large genomes. Here we investigate the range of nuclear DNA contents across which the standard AFLP protocol can be used. METHODS: AFLP fingerprinting was conducted on an automated platform using the standard protocol (with 3 + 3 selective bases) in which DNA fragments are visualized as bands. Species with nuclear DNA contents ranging from 1C = 0.2 to 32.35 pg were included, and the total number of bands and the number of polymorphic bands were counted. For the species with the smallest C-value (Bixa orellana) and for one of the species with a large C-value (Damasonium alisma), alternative protocols using 2 + 3 and 3 + 4 selective bases, respectively, were also used. KEY RESULTS: Acceptable AFLP traces were obtained using the standard protocol with 1C-values of 0.30-8.43 pg. Below this range, the quality was improved by using 2 + 3 selective bases. Above this range, the traces were generally characterized by a few strongly amplifying bands and noisy baselines. Damasonium alisma, however, gave more even traces, probably due to it being a tetraploid. CONCLUSIONS: We propose that for known polyploids, genome size is a more useful indicator than the 1C-value in deciding which AFLP protocol to use. Thus, knowledge of ploidy (allowing estimation of genome size) and C-value are both important. For small genomes, the number of interpretable bands can be increased by decreasing the number of selective bases. For larger genomes, increasing the number of bases does not necessarily decrease the number of bands as predicted. The presence of a small number of strongly amplifying bands is likely to be linked to the presence of repetitive DNA sequences in high copy number in taxa with large genomes.

Cell Nucleus↗
PubMed2003

AFLPinSilico, simulating AFLP fingerprints.

SUMMARY: A drawback of the Amplified Fragment Length Polymorphism (AFLP) fingerprinting method is the difficulty to correlate the different fragments with their DNA sequence. The AFLPinSilico application presented here simulates AFLP experiments run on either cDNA or genomic sequences, producing virtual fingerprints that allow high throughput identification of AFLP fragments. The program also enables biologists to manage experiments through simulations done beforehand, thereby reducing the number of experiments that have to be run. AFLPinSilico is available through the www or as a stand-alone version, through a command line executable (available upon request, for any platform running PERL).

Algorithms↗
PubMed1996

Prediction of heterosis from DNA fingerprints in chickens.

To assess the value of DNA fingerprints for the prediction of heterosis in chickens, retrospective analyses of data from three crossbreeding experiments and DNA fingerprints (DEP) of parental strains were conducted using two minisatellite and one middle-repetitive DNA probes. DEP bands were assessed on pooled DNA samples of 10-15 individuals per parental genetic group. The number of DEP bands evaluated in the experiments ranged from 81 to 139. The probes varied in their predictive value, but predictability of heterosis generally increased with multiple probes. Highly significant correlations (0.68-0.87) between band sharing ratios (SH) and heterosis were found in 25 crosses of White Leghorns in the first egg production cycle for age at sexual maturity, egg production, and mature body weight: traits with heterosis of 10% or more of the means. Regressions on SH explained 78.4% of the variation in heterosis in age at sexual maturity, 60.2% in egg production and 46.4% in mature body weight. For "broiler" traits with heterosis of < 1%, none of the correlations, based on 13 crosses, were significant. It was concluded that multilocus probe DFP of pooled DNA samples show promise as predictors of heterosis.

Animals↗
PubMedDate not supplied

Evaluation of genetic variation within and between different chicken lines by DNA fingerprinting.

A DNA fingerprinting technique was used to assess the extent of genetic variation within and between 12 lines of chickens of different origin. DNA fingerprints were obtained with the restriction endonucleases AluI and HinfI and with the oligonucleotide probes (CA)8, (CAC)5, (GGAT)4, and (GACA)4. Different methods that are based on band sharing and band frequency were tested to estimate genetic variation within and between populations. The genetic variability was significantly different within the populations. It was highest in a commercial broiler line and lowest in a White Leghorn inbred line. All three measures of genetic variation within populations provided results that were highly correlated (r = 0.89-0.97). A lower degree of correspondence was observed for the estimation of genetic variation between populations by three different methods. This is shown by differences of the phylogenetic trees and by coefficients of correlation between these measures ranging from r = 0.51-0.96.

Animals↗
PubMedDate not supplied

Analytical aspects of population-specific DNA fingerprinting for individuals.

An emerging problem of some interest is whether we can determine the population membership of a single individual, using a population-specific "genetic fingerprint." The levels of accuracy and precision required are beyond the reach of allozyme analysis, and attention has shifted to DNA polymorphisms. There are different types of DNA markers available for population surveys: RFLPs, mini- and micro-satellites, and RAPDs, and each type has its own strengths and weaknesses. We present a generic analysis that relates gene pool separation to our ability to assign individuals, an analysis that does not depend on the type of marker. We then review strengths and weaknesses of different DNA markers, in the context of DNA fingerprinting. Codominant loci are best. It is possible to gain more information per marker with multiallelic loci, but diminishing returns set in rapidly, and it is better to add loci. A modest number of independent loci is best, each locus with a modest number of alleles and with each allele in modest frequency.

DNA Fingerprinting↗
PubMed1995

AFLP: a new technique for DNA fingerprinting.

A novel DNA fingerprinting technique called AFLP is described. The AFLP technique is based on the selective PCR amplification of restriction fragments from a total digest of genomic DNA. The technique involves three steps: (i) restriction of the DNA and ligation of oligonucleotide adapters, (ii) selective amplification of sets of restriction fragments, and (iii) gel analysis of the amplified fragments. PCR amplification of restriction fragments is achieved by using the adapter and restriction site sequence as target sites for primer annealing. The selective amplification is achieved by the use of primers that extend into the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotides flanking the restriction sites. Using this method, sets of restriction fragments may be visualized by PCR without knowledge of nucleotide sequence. The method allows the specific co-amplification of high numbers of restriction fragments. The number of fragments that can be analyzed simultaneously, however, is dependent on the resolution of the detection system. Typically 50-100 restriction fragments are amplified and detected on denaturing polyacrylamide gels. The AFLP technique provides a novel and very powerful DNA fingerprinting technique for DNAs of any origin or complexity.

Base Sequence↗
PubMed1998

Analysis of tuberculosis transmission between nationalities in the Netherlands in the period 1993-1995 using DNA fingerprinting.

Immigration from high prevalence areas may contribute to an increased risk of tuberculosis in Europe. This study aimed at quantifying transmission of tuberculosis between and within nationalities among residents of the Netherlands. DNA "fingerprints," on the basis of restriction fragment length polymorphism using marker IS6110, were made of all Mycobacterium tuberculosis isolates in the Netherlands from January 1993 through June 1995. Clusters were defined as groups of patients that had isolates with identical fingerprints. It was assumed that the probability of a patient being the source of a cluster was proportional to the incidence rate of potential sources times the probability that a potential source would give rise to a cluster. The transmission index was defined as the average number of secondary cases of infectious tuberculosis caused directly or indirectly through recent transmission by a single potential source case and was used to estimate the effective reproductive rate associated with recent transmission, ReFAST. Among a total of 623 Dutch tuberculosis cases, 17% (95% confidence interval 9-25%) of cases were attributable to recent transmission from a non-Dutch source. The transmission index varied strongly by nationality, and was highest among the Surinamese (1.3), Moroccan (0.8), and Turkish (0.8) populations; ReFAST was 0.26. Aggregation of tuberculosis cases of given nationalities within clusters was most pronounced among recent immigrants from Somalia and (ex-)Yugoslavia. The authors conclude that differences in transmission between subpopulations can be quantified and may be used to evaluate and direct tuberculosis control.

Adolescent↗