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Analysis of population genetic structure by DNA fingerprinting.

DNA fingerprint similarity is now being used widely to make inferences about the genetic structure of natural and domesticated populations, often with little regard to the limitations of such data. This paper provides an overview of the statistical theory of DNA fingerprint analysis with special focus on applications to natural populations for which little if anything is known about the detailed genetics of the DNA profiles. Approaches to estimating individual and population homozygosity, effective population size, population subdivision, and relatedness are reviewed, and issues concerning the biases and sampling properties of the statistics are discussed.

DNA Fingerprinting

Genetic variation between strains of the Mediterranean fruit fly, Ceratitis capitata, detected by DNA fingerprinting.

DNA fingerprinting has been used to detect genetic variation in the Mediterranean fruit fly, Ceratitis capitata. Three different probes have been identified that can be used to detect DNA restriction fragment length polymorphisms between strains of this species. The strains used in this study differ only in terms of their geographic origin or genetic background. One of the probes used is the bacteriophage vector M13, and the other two are repetitive sequences derived from the medfly genome based on a weak homology to M13. Within a strain, each probe produces a consistent restriction fragment profile that is not affected by the method or timing of DNA extraction. Between strains, when M13 is used as a probe, an average of 10% of the observable bands are polymorphic. Use of the medfly genomic sequences as a probe increases the proportion of polymorphic bands between strains up to 30%. The fact that genetic differences between even such closely related strains can be reliably detected by this method holds great promise for studies of insect pests including the ability to monitor the movements of pest species, determining the extent of genetic variation in pest populations, and in making identifications from otherwise unidentifiable material.

Animals

"Sexing" deoxyribonucleic acid (DNA) on DNA fingerprint gel: an internal control for DNA fingerprint evidence.

Deoxyribonucleic acid (DNA) isolated from male and female fresh blood samples was processed exactly as for routine DNA fingerprint analysis; that is, the DNA was digested with particular restriction endonucleases and fractionated by agarose gel electrophoresis. Ultraviolet (UV) visualization of ethidium-bromide (EtBr)-stained gels revealed a sex-specific banding pattern, which depended only on the restriction enzyme used. By means of this test, which is based on direct detection of particular sex-specific restriction fragments in human DNA digests, the authors succeeded in determining the sex of DNA obtained from biological specimens recovered as criminal evidence in rape cases. The data obtained demonstrate that direct sexing of DNA on DNA fingerprint gel appears to be useful as an intermediate control step in DNA fingerprinting analysis used for the purpose of assailant identification.

DNA

Re-evaluation of hydatidiform mole by DNA fingerprint method: the discrepancy in the diagnoses by pathological finding and the DNA fingerprint method.

Ten cases of hydatidiform mole (HM) were analysed by the DNA fingerprint method. DNA samples were prepared from HM tissue of HM and maternal and paternal bloods, followed by digestion with HaeIII restriction endonuclease, applied to agarose gel, and then transferred to a nitrocellulose filter. The filter was hybridized using mini-satellite DNA as a probe. A case of partial HM diagnosed by histological findings was revealed to be complete HM using DNA fingerprinting. The case was suggested to be a twin pregnancy with a cHM and an abortion, because the DNA fingerprint of hydropic tissue showed a paternal pattern and that of the non-hydropic tissue did not show such a pattern. DNA fingerprinting was considered to be useful in distinguishing complete HM from partial HM.

Adult

Probing identity: the changing face of DNA fingerprinting.

DNA-fingerprinting technology has made a very rapid transition from being a research laboratory discovery to an applied science widely understood by, and of interest to, the general public. However, DNA fingerprinting is often portrayed as being a single generic technology, rather than a complex evolving mixture of methodologies, where specific applications demand selection of appropriate probes and techniques.

Animals

The similarity index and DNA fingerprinting.

DNA-fingerprint similarity is being used increasingly to make inferences about levels of genetic variation within and between natural populations. It is shown that the similarity index--the average fraction of shared restriction fragments--provides upwardly biased estimates of population homozygosity but nearly unbiased estimates of the average identity-in-state for random pairs of individuals. A method is suggested for partitioning the DNA-fingerprint dissimilarity into within- and between-population components. Some simple expressions are given for the sampling variances of these estimators.

Alleles

Detection of somatic mutations in tumours of diverse types by DNA fingerprinting with M13 phage DNA.

Hybridization of M13 phage DNA to Southern blots of human DNA produces an individual-specific DNA fingerprint. In this study, tumour and lymphocyte DNA from a series of patients with melanoma, Merkel-cell carcinoma, Burkitt's lymphoma and Wilms' tumour was probed with M13 DNA to detect somatic mutations in the DNA of the tumours. Somatic changes were observed in tumour DNA of 16 out of the 28 cases examined. This frequency compared favourably with the frequency with which tumour-specific changes have been found when using the Jeffreys DNA fingerprinting probe 33.15, and demonstrates that M13 DNA provides a useful additional probe for the study of somatic changes in tumours. The finding that multiple DNA fragments were lost or gained in DNA fingerprints from individual tumours indicates a marked degree of complexity in the genetic changes involved in the evolution of certain human cancers.

Bacteriophages

Detection of genomic alterations in carcinogen-induced mouse liver tumors by DNA fingerprint analysis.

DNA fingerprint analysis was used to study structural abnormalities in the genome of mouse liver tumor cells. Liver tumors were induced in three strains of mice, namely C57BL/6J, C3H/He and B6C3F1, by a single injection of 20 micrograms/g body wt. diethylnitrosamine on day 15 after birth. DNA from liver tumors was digested with Hinfl restriction enzyme and hybridized on Southern blots with wild-type bacteriophage M13 DNA as probe. The resulting fingerprints of tumor DNA were compared with those of DNA from normal liver tissue. Genomic aberrations were detected in two out of 68 tumors analyzed, one stemming from a C57BL/6J and the other from a C3H/He mouse.

Animals

Characterization of the sequence of colonization and nosocomial candidemia using DNA fingerprinting and a DNA probe.

The objective of this hospital-based study was to determine the relationship between colonizing and infecting strains of Candida species and Torulopsis glabrata. Surveillance cultures from high-risk patients were paired with subsequent bloodstream isolates. Organisms were typed by using restriction endonuclease digestion of chromosomal DNA with BstNI and EcoRI, followed by Southern hybridization with a DNA probe (pBD4) derived from Saccharomyces cerevisiae. Sixteen patients for whom documented colonization preceded documented bloodstream infection were identified. The mean time between obtainment of surveillance isolates and obtainment of bloodstream isolates was 8 days, with a range of 1 to 423 days. For 15 (94%) of 16 patients, the DNA fingerprint pattern (using BstNI) of the surveillance isolate was identical to that of the bloodstream isolate. Isolates from 13 (81%) of 16 patients were unique to those patients. Typing by Southern hybridization with the pBD4 probe was less discriminating. We conclude that for a well-defined subset of hospitalized patients who were colonized by Candida species before developing nosocomial candidemia, the colonizing and infecting strains were identical, suggesting endogenous acquisition of infection. Restriction endonuclease digestion of chromosomal DNA was shown to be a discriminating and reproducible typing method for Candida species and T. glabrata.

Blotting, Southern

Detection of DNA alterations in human bladder tumors by DNA fingerprint analyses.

DNA fingerprint analyses were used to examine the constitutional and tumor DNA from 22 bladder tumor patients. DNA alterations, such as loss of bands, new bands, and intensity shifts were observed in 10 of the 22 patients. The most frequent DNA alteration, occurring in 80% of the patients, was a complete loss of one or several bands. Fingerprint abnormalities were present both in low-malignant superficial tumors and in high-malignant invasive tumors, but were also lacking in the latter group. Apparently no relationship exists between fingerprint abnormalities and gross chromosomal aberrations or the proportion of S-phase cells as measured by flow cytometry or development of recurrent tumors during a limited observation period. Thus, whether fingerprint aberrations express genetic alterations directly involved in the malignancy potential of bladder carcinoma remains an open question.

Adult

Genetic factors accountable for line-specific DNA fingerprint bands in quail.

DNA fingerprints, prepared from mixes of DNA of individuals sampled from lines of Japanese quail selected for high or low 4-week body weight, were used to evaluate the relative contribution of several evolutionary forces to genetic diversity among populations. Comparisons between lines--two replicates of each selection direction and a control unselected line--were used to determine the frequency of line-specific DNA fingerprint bands produced by each of three major evolutionary forces: 1) mutation; 2) genetic drift; 3) selection. The latter force is expected to generate line-specific bands only if there is linkage disequilibrium between DNA fingerprint loci and quantitative loci (QTLs) controlling body weight. Using probes 33.6 and R18.1, an average of 48.4 DNA fingerprint bands in each line were analyzed. On average, 27.8 bands were found to be line-specific among the 96.8 (2 x 48.4) bands analyzed in an average comparison between pairs of lines. Based on the frequencies of line-specific bands in each particular comparison, it was calculated that 21% of the line-specific bands were due to mutation, 11% due to a single genetic drift event, 11% due to selection, 21% due to the combined effects of genetic drift and selection, 22% due to double independent events of genetic drift, and 14% due to undefined factors. Although evidence was found for a high frequency of genetic changes attributable to genetic drift, and a higher than expected frequency of linkage disequilibrium, the emphasis of this report is on the methodology suggested rather than on the particular results.

Animals

Does chemotherapy of hematological malignancies affect DNA fingerprint pattern?

We analyzed DNA fingerprints of lymphoma patients to find out whether DNA damage caused by irradiation and chemotherapy can result in DNA fingerprint changes, and whether the differences found previously in leukemia patients could be partially due to the treatment. In this study we did not find any post-treatment DNA fingerprint differences in 33 lymphoma patients, concluding, that the therapy of hematological malignancies does not affect DNA fingerprint patterns. Further, the variations of methylation do not either explain the detected differences in leukemic patients.

DNA Damage

Sizing bands on autoradiograms: a study of precision for scoring DNA fingerprints.

We replicated DNA fingerprints of snapping turtles (Chelydra serpentina) and hypervariable restriction fragments of red-winged blackbirds (Agelaius phoeniceus) to estimate the between-blot and between-lane components of variance in molecular weights of restriction fragments. Molecular weight standards were included in every lane, and bands were sized using a sonic digitizer. In both studies, a strong positive correlation was found between band size and coefficient of variation (CV; mean = 0.7%). In the DNA fingerprint study, 26% of the variance in estimates of band size was due to differences between blots, 10% due to differences between lanes on the same blot, and 64% due to error in the digitizing process. In the restriction fragment length polymorphism (RFLP) study, 16% of the variance was due to difference between lanes, and 84% to digitizing. Statistical models were developed to measure the effect of sizing error on identifying identical fragments in different lanes or on different blots, in categorizing distinct alleles, and in determining the size of bins in operational allele definitions. We suggest that the distance between bands be at least 2.8 standard deviations (SD) before they are declared different at alpha = 0.05, and 3.7 SD for alpha = 0.01. A variation in CVs strongly indicates that empirical relationships between SD and band size must be used to decide if two bands represent the same allele. Alleles must be at least 3.9 SD apart before the chance of assigning new observations in error falls below 0.05. We suggest that a minimum bin width of 16 SD is necessary before the chances of assigning a band to the wrong bin falls below 0.05.

Alleles

Improved resolution and sensitivity of human DNA fingerprinting by specific-primed labelling of M13 DNA.

A method to label M13 DNA probe by primer extension using a specific oligonucleotide primer is described. The method specifically labels the two 15-bp repeats in M13 DNA which hybridize to target DNA giving rise to DNA fingerprinting patterns. The M13 probe labelled by this method gave superior DNA fingerprinting patterns that that labelled by random primers. As little as 0.25 microgram of target DNA was sufficient for DNA fingerprinting. Non-isotopic labelling by the specific primer also showed improved DNA fingerprinting pattern. The results demonstrate the methodology to improve DNA fingerprinting based on M13 DNA probe.

Base Sequence

Principles and recent advances in human DNA fingerprinting.

Since 1985, DNA typing systems have played an increasingly important role in many aspects of human genetics, most notably in forensic and legal medicine. This article reviews the development of multilocus and single locus minisatellite DNA probes, and more recently the use of PCR to amplify hypervariable DNA loci, as well as discussing the biological properties of the unstable regions of DNA which form the basis of almost all DNA fingerprinting systems.

Base Sequence

[Analysis of genetic distances between populations using human DNA "fingerprints" detected by a phage M13 DNA probe].

The frequencies of different electrophoretic bands in DNA fingerprints detected by phage M13 DNA probe in two populations of the Kirov district were determined. The DNA polymorphisms observed in these two populations were compared with each other and with those of the Krasnodar populations, and pseudogenetic distances were calculated. The mean genetic distance between two Kirov populations was 0.072, this between every Kirov and Krasnodar population being 0.21 and 0.22.

Bacteriophages

RAPD analysis of Campylobacter isolates: DNA fingerprinting without the need to purify DNA.

A method was developed to obtain reproducible DNA fingerprints from Campylobacter by PCR-based amplification, without the need to isolate total DNA. Randomly amplified polymorphic DNA (RAPD) profiles were generated with three randomly designed 10-mers, using each separately as an amplification primer. A range of C. jejuni serotypes could be typed by RAPD analysis. Depending on the primer, the analysis of RAPD profiles resulted in different levels of discrimination between the strains. Clear correlations were observed between results of RAPD analysis and serotyping. Two of the primers tested generated RAPD profiles which allowed discrimination of strains within given Penner and Lior serotypes.

Base Sequence