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PubMed1992

Genomic variations in mosquitocidal strains of Bacillus sphaericus detected by M13 DNA fingerprinting.

The genomic variation of Bacillus sphaericus reference and local strains belonging to different serotypes was examined by DNA fingerprinting. A phage M13 DNA probe detected a number of variable fragments in the restriction digests of total strain DNAs. The patterns of band distribution showed a certain homology among mosquitocidal strains, expressed by similarity index D and might be a reliable criterion for assessing the level of genomic similarity between closely related strains. An important advantage of DNA fingerprinting is the differentiation of one bacterial strain from another, both expressing common phenotype and possessing highly similar genomic portions. The strain variation revealed by the M13 probe will be useful for characterization of individual strains within a serotype. It could help as well to solve some uncertain cases based on the results obtained by other methods of identification.

Animals↗
PubMed2005

DNA fingerprinting on trial: the dramatic early history of a new forensic technique.

The early history of "DNA fingerprinting" in the UK might have been different were it not for the accounts of two dramatic courtroom trials, made by the participants and the media, in the mid-1980s. But these reports, which misrepresented the importance DNA evidence had in the trials, left a strong impression on the British public and on judges on both sides of the Atlantic. These trials, widely considered to be the first "victories" for DNA fingerprinting, have been frequently cited as proof of the utility and reliability of the technique, in both the UK and beyond. But in reality, it was the threat of DNA evidence being used rather than the integrity or validity of it that resolved these cases. At that time, DNA fingerprinting was still in its infancy, an untried and untested technology.

Criminal Law↗
PubMed2001

Identifying the origin of single corneal cells by DNA fingerprinting: part II-- application to limbal allografting.

PURPOSE: Successful limbal allotransplantation allows regression of limbal stem cell deficiency features. Transplant survival is presumed if clinical improvement occurs. However, positive proof of surviving transplanted stem cells remains difficult. This follow-up study attempted to prove donor cell survival 5 years after limbal stem cell allograft in one woman with aniridia. METHODS: Impression cytology and single-cell DNA fingerprinting were used to investigate a previously studied patient. Corneal epithelial cells were harvested from five sites and isolated by micromanipulation. Polymerase chain reaction and short tandem repeat profiling were used to obtain forensic standard "DNA fingerprints" from single cells. (The technique is described in the preceding article, Part I.) Blood samples yielded host and donor DNA for comparison. Negative controls were performed for impression cytology and polymerase chain reaction. Simultaneous micro-scrape samples were also taken. RESULTS: Impression cytology samples permitted informative DNA fingerprints from all corneal sites and represented 76% (23/30) of tested cells. Fifty percent (15/30) of the fingerprints were "specific" but 83% (19/23) matched the host DNA fingerprint. The remaining 17% (4/23) represented contamination from various sources. Specific fingerprints were obtained in 55% (10/18) of the cells from micro-scrape samples. All samples giving sufficient information matched the host DNA fingerprint. All tested blood samples gave specific fingerprints. None of the sampled corneal cells gave a donor DNA fingerprint. CONCLUSIONS: In a single patient, no detectable long-term donor cell survival exists at 5 years. Positive identification would have provided unequivocal proof of donor cell survival. This technique gives useful information even if contamination occurs.

Aniridia↗
PubMed1992

Likelihoods of multilocus DNA fingerprints in extended families.

A concept for the application of complex pedigree analysis to multilocus DNA fingerprinting is described. By following this approach, the extent to which the DNA fingerprints of grandparents influence the phenotype likelihoods of their offspring was determined. It was demonstrated by simulation that approximately 90% of paternity disputes can be solved if mother, child, and paternal grandparents, instead of the putative father, are tested. If only phenotype information on a single paternal sib is allowed for, true paternity will be detected with reasonable persuasive power in up to 64% of cases. Exclusion of false paternity remains possible for 40% of cases. Finally, the analysis concept is modified by reducing the number of genotype variations considered in likelihood computations. This time-saving procedure is shown to yield sufficiently accurate likelihoods in the analysis of both simulation data and multilocus DNA fingerprints obtained in two large families.

DNA Fingerprinting↗
PubMed2004

Demonstration of a gastric bioptic specimen mix-up by laser capture microdissection (LCM) and DNA fingerprinting.

We demonstrate here the successful use of laser capture microdissection (LCM) and DNA fingerprinting in the identification of a case of gastric bioptic specimen mix-up. A 70-year-old man, suffering from chronic atrophic gastritis, underwent to a gastric biopsy and received a diagnosis of gastric cancer. In the absence of any clinical evidence of gastric cancer, a specimen mix-up was suspected. LCM was used to retrieve gastric cells from the histologic slide, classified as gastric carcinoma, and suspected to be mislabelled. DNA was extracted from microdissected cells, and a total of 16 different genetic loci were analyzed, using an identity test. Comparison of the results with those obtained using DNA extracted from a control slide, and from patient's saliva, demonstrated a distinct DNA fingerprint pattern in all genetic markers examined, clearly indicating the occurrence of a specimen mix-up. The combined use of LCM and DNA fingerprinting represents the most accurate and sophisticated method available for the identification of specimen mix-up, especially when only the tissue on the suspected slide is available.

Aged↗
PubMed1991

DNA fingerprinting of Streptococcus uberis based on polymorphism of DNA encoding rRNA.

The rRNA gene restriction patterns of strains of Streptococcus uberis and Strep. parauberis were determined. Chromosomal DNA was digested with endonucleases and probed with radiolabelled DNA complementary to rRNA synthesized by random oligonucleotide priming using reverse transcriptase. Reproducible restriction patterns were obtained which readily distinguished Strep. uberis and Strep. parauberis. In addition, considerable variations in the patterns at the intra-specific level were observed indicating that rRNA gene restriction profiles are of epidemiological, as well as taxonomic value.

Blotting, Southern↗
PubMed2001

Quantifying mosquito biting patterns on humans by DNA fingerprinting of bloodmeals.

A major debate in infectious disease epidemiology concerns the relative importance of exposure and host factors, such as sex and acquired immunity, in determining observed age patterns of parasitic infection in endemic communities. Nonhomogeneous contact between hosts and vectors is also expected to increase the reproductive rate, and hence transmission, of mosquito-borne infections. Resolution of these questions for human parasitic diseases has been frustrated by the lack of a quantitative tool for quantifying the exposure rate of people in communities. Here, we show that the polymerase chain reaction (PCR) technique for amplifying and fingerprinting human DNA from mosquito bloodmeals can address this problem for mosquito-borne diseases. Analysis of parallel human and mosquito (resting Culex quinquefasciatus) samples from the same households in an urban endemic focus for bancroftian filariasis in South India demonstrates that a 9-locus radioactive short-tandem repeat system is able to identify the source of human DNA within the bloodmeals of nearly 80% of mosquitoes. The results show that a person's exposure rate, and hence the age and sex patterns of exposure to bites in an endemic community, can be successfully quantified by this method. Out of 276 bloodmeal PCR fingerprints, we also found that on average, 27% of the mosquitoes caught resting within individual households had fed on people outside the household. Additionally, 13% of mosquitoes biting within households contained blood from at least 2 people, with the rate of multiple feeding depending on the density of humans in the household. These complex vector feeding behaviors may partly account for the discrepancies in estimates of the infection rates of mosquito-borne diseases calculated parasitologically and entomologically, and they underline the potential of this tool for investigating the transmission dynamics of infection.

Adolescent↗
PubMed1991

DNA fingerprinting in the detection of residual disease in acute leukemia.

DNA-fingerprint (DNA-F) analysis of leukemic cells can reveal new restriction fragments in some patients with acute leukemia. These genomic alterations can be used as molecular markers of residual disease. In this study, the sensitivity and clinical use of DNA-F analysis was evaluated. The detection limit for minority DNA was studied in six cell dilution series. An increasing ratio (1-20%) of cells was mixed with cells from another individual; intense extra bands could be identified in the DNA extracted from cell mixtures containing 5% of minority cells. In clinical samples from five of 19 ALL and six of 18 AML cases, the DNA-F of blast cells had new bands which could be followed during remission; in two cases the extra bands were intense enough to be detected at less than 5%. Studies of 37 patients with leukemia demonstrated that DNA-F analysis provides an additional tool for the detection of residual disease in acute leukemia.

Acute Disease↗
PubMed1992

Evidence of transmission of tuberculosis by DNA fingerprinting.

OBJECTIVE: To determine whether a subject who had died of tuberculous meningitis had been infected by a neighbour. DESIGN: Retrospective comparison of isolates of Mycobacterium tuberculosis from the two cases and from 10 controls by DNA fingerprinting. SETTING: Public Health Service Reference Laboratory for Mycobacteria and bacterial molecular genetics unit of the London School of Hygiene and Tropical Medicine. SUBJECTS: Deceased and neighbour; 10 controls from the same city, from whom isolates had been collected over three months before the subject's death. MAIN OUTCOME MEASURES: Identity and similarity values (SAB) between fingerprint patterns from different isolates obtained by hybridisation of restriction fragments produced by PvuII with a probe from the insertion element IS6110/986, present in multiple copies throughout the genome of M tuberculosis. RESULTS: Isolates from the two cases under investigation had identical fingerprints whereas those from the controls were all distinct. Two clusters of isolates with a similarity coefficient > 0.25 were identified: in one, four out of five patients were born in the midlands (the birth place of the fifth was not known) and in the other all three patients were born in the Indian subcontinent. CONCLUSIONS: The data are consistent with, but do not prove, transmission of tuberculosis from the neighbour to the deceased. Geographical separation of the pools of infection may have led to the evolution of distinct clusters of fingerprint patterns. DNA fingerprinting of M tuberculosis is a powerful new tool for study of the epidemiology and pathogenesis of tuberculosis.

DNA Fingerprinting↗
PubMed1999

Abnormal patterns of immunoglobulin heavy chain gene DNA fingerprinting during chronic phase chronic myeloid leukemia.

Using IgH DNA fingerprinting we have previously demonstrated clonal immunoglobulin heavy chain (IgH) gene rearrangements during chronic phase (CP) chronic myeloid leukemia (CML) in patients destined to develop lymphoid blast crisis (L-BC). In view of this we decided to follow a cohort of CP CML patients to determine the frequency with which abnormal IgH fingerprints are found and their relationship, if any, to treatment regimen. Thirty three, initially CP, CML patients were studied on 111 occasions over a 16 month period using consensus PCR amplification of the third complementarity determining region (CDR3) of the IgH gene and high resolution polyacrylamide gel electrophoresis (IgH DNA fingerprinting). Of these 33 patients, thirteen received interferon-alpha (IFN) containing regimens and 15 non-IFN containing regimens throughout the study period. Five patients received variable therapy. During the period of observation 7 patients experienced disease progression: 5 accelerated phase, I L-BC and I myeloid blast crisis (M-BC). Abnormal IgH fingerprints were seen in 29 of the 111 (26%) specimens analysed. The 28 patients who received uniform therapy (IFN or non-IFN) over the 16 months were classified as "normal" (n = 18, normal pattern on all occasions) or "abnormal" (n = 10, abnormal on 1 or more occasions). Analysis by patient group (normal vs abnormal) showed that fingerprint abnormalities were associated with an elevated peripheral blood lymphocyte count (p = 0.0001) but not with changes in the total white cell count. Comparison of the IFN vs. non-IFN groups showed the former all had normal patterns whereas 10 of 15 non-IFN therapy patients were abnormal (p = 0.00023). The peripheral blood lymphocyte counts in the normal vs abnormal patients within the non-IFN group were not significantly different. The patient who developed L-BC demonstrated a persistent IgH fingerprint pattern abnormality from 7 months prior to the diagnosis of L-BC. The M-BC patient had a normal pattern at all times. We conclude that: (1) abnormal IgH fingerprints are found in a significant number of CP CML patients; (2) in this cohort the use of IFN was associated with normal CP CML IgH fingerprints, and (3) detection of abnormal IgH fingerprints may be highly predictive for the lineage of impending blast crisis.

DNA Fingerprinting↗
PubMed1992

DNA fingerprinting of Chlamydia trachomatis by use of ribosomal RNA, oligonucleotide and randomly cloned DNA probes.

DNA fingerprinting of 15 reference strains and 24 clinical isolates of Chlamydia trachomatis, 2 strains of C. psittaci and one strain of C. pneumoniae was studied by use of universal 16 + 23S RNA from Escherichia coli, 16S rDNA-directed oligonucleotide and randomly cloned chlamydial DNA probes. The rRNA-gene restriction patterns (ribotypes) enabled the differentiation of chlamydial species. Following DNA cleavage by restriction endonuclease PvuII, lymphogranuloma venereum and trachoma biovars of C. trachomatis could be differentiated. An oligonucleotide, designed to hybridize the C. trachomatis 16S rDNA, also allowed for both species-specific identification and biovar typing of C. trachomatis human strains. Molecular typing system using 3 lambda clones containing C. trachomatis serotype E random DNA inserts, combined to ribotyping, revealed 12 groups of variable banding patterns within C. trachomatis, and could provide an alternative epidemiological tool.

Autoradiography↗
PubMed1991

Detection of somatic DNA alterations in ovarian cancer by DNA fingerprint analysis.

The M13 phage single-strand DNA probe which recognizes highly polymorphic loci was applied to HinfI-digested DNA isolated from tumor tissue and peripheral leukocytes from 20 patients with ovarian cancer. An average of 22 minisatellite-containing DNA fragments were observed per individual. DNA fingerprint analysis revealed a change in restriction-fragment-length patterns in the DNA from 12 of 20 (60%) tumors compared with the patient's constitutional DNA. Deletion of one or more bands from the tumor was recognized by the probe in seven cases, new bands were identified in two, and intensity shift was demonstrated in eight. The authors conclude that the unmapped M13 minisatellite probe is a useful method for identifying cancer-related somatic DNA alterations.

Adult↗
PubMed1995

DNA fingerprinting in captive population of the endangered Siberian crane (Grus leucogeranus).

DNA fingerprinting was used to estimate genetic diversity within the endangered Siberian crane (Grus leucogeranus) captive population consisting of several dozens of founders originating from the two wild populations of eastern and western Siberia. Similarity and difference among captive individuals were demonstrated by the unweighted pair-group (UPGMA) clustering procedure. Quantitative characteristics of the eastern and western captive population groups such as average percentage differences (APD) and heterozygosity showed a high extent of genetic variability of 77.9-79.3% and heterozygosity of 0.85-0.72 within each group. Genetic heterogeneity of the captive population structure observed here provides guidelines for management of the species gene pool in captivity. These data also indicate that monitoring of genetic diversity through DNA fingerprinting can facilitate the efforts of Siberian crane management and restoration.

Animals↗
PubMed1991

DNA fingerprinting in cattle using oligonucleotide probes.

Oligonucleotide probes specific for simple tandem repeat sequences produce individual specific DNA fingerprints in man and all animal species tested so far. Here 11 different synthetic probes were hybridized to bovine genomic DNAs which had been digested with the restriction endonucleases HinfI, AluI and HaeIII. Two of these probes gave DNA fingerprint patterns which were analysed for three German breeds. Different parameters were calculated, such as the average number of bands per individual or the probability of finding identical fingerprints in two unrelated individuals. The number of polymorphic bands varies from 11 to 23 in the different breeds and the probability of finding the same banding pattern in two unrelated individuals ranges from 1.5 x 10(-7) to 2.4 x 10(-7). Hence this DNA fingerprinting procedure allows precise identification of individuals. It is also a useful additional method for paternity testing in cattle.

Animals↗
PubMed2003

DNA fingerprinting of mosses.

Our study introduces the use of DNA fingerprinting of clonal plants in combination with phylogenetic and vegetation studies as a prospective forensic tool in criminal investigations. In this homicide case, the bryophyte species found on the suspects were identified as Brachythecium albicans, Calliergonella lindbergii, and Ceratodon purpureus. Colonies of all three species occurred at the crime site. DNA fingerprinting analyses were conducted for B. albicans and C. lindbergii, which were expected to reproduce mainly clonally, unlike C. purpureus, and included samples found on the suspects and samples collected from the crime site and other locations. It was concluded that B. albicans found on the suspects was likely to originate from the crime scene and that the sample of C. lindbergii may also have originated from the same site.

Adult↗
PubMed2003

Statistical evaluation of bacterial source tracking data obtained by rep-PCR DNA fingerprinting of Escherichia coli.

Pattern recognition has been applied to environmental systems for identification of numerous pollution sources including aerosolized lead and petroleum hydrocarbons. In recent years, DNA fingerprinting has gained widespread application as a means to characterize genetic variations for such purposes as microbial source tracking. This approach, however, is strongly dependent on the statistical and image analyses applied. Several statistical analyses of rep-PCR DNA fingerprints were assessed as a means to differentiate between potential sources of fecal contamination. GelCompar II and methods based on penalized discriminant analysis (PDA) and k-nearest neighbors (KNN) classification procedures were used to differentiate between 10 source groups within a library containing DNA fingerprints of 548 Escherichia coli isolates from known human and nonhuman sources. KNN performed significantly better than PDA in a jackknife analysis, though the library was not large enough to detect significant differences between GelCompar II and the other two methods. GelCompar II and KNN both attained > or = 90% correct classification in a holdout procedure. In addition, interpoint distance analyses indicate coherency within source groups, while library randomization demonstrated that KNN does not create artificial groupings. This investigation stresses the need to understand limitations of statistical analyses used in pattern recognition of DNA fingerprints.

DNA Fingerprinting↗
PubMed1994

DNA fingerprinting and the substantive law.

The ability to analyse human DNA has the potential to revolutionize legal proceedings--both civil and criminal--especially those which turn on biological or genetic evidence. Examining the positive and negative aspects of DNA fingerprinting, it would appear that the benefits certainly outweigh any disadvantages. In paternity determinations, DNA fingerprinting renders otiose many of the controversial concerns, particularly regarding the determination of legal probability, providing as it does unequivocal evidence of blood relationship. Furthermore, in criminal law, its value is well-documented. The relevant legislation is already in place, therefore no further legislation is required to enable South African law enforcement agencies to make use of the DNA test. The apparently high cost may, however, frustrate its general application to the civil law but with the assistance of the state and law societies, the envisaged problem could be alleviated. To date no other forensic tests rival the accuracy of the DNA test.

Cost-Benefit Analysis↗