2. A Multilocus DNA Fingerprint with Built-in Security Devices

1994 ◽  
Vol 34 (3) ◽  
pp. 256-262
Author(s):  
Michael Krawczak ◽  
Jörg Schmidtke ◽  
Jörg T Epplen ◽  
Ingo Hansmann ◽  
Ulrike Thies

An unusual case of paternity testing is reported in which determination of paternity was an essential part of a genetic diagnosis. A.Y-chromosomal abnormality, observed in a 33-year-old male whose wife had experienced a series of spontaneous abortions, was not found in his alleged father. DNA fingerprinting with the oligonucleotide multilocus probe (CAC)5 yielded two aberrant bands for the proband, i.e. bands exhibited by neither parent. This finding resulted in a comparatively low paternity probability of 0.02934 which is suggestive of, but does not unequivocally prove, false paternity. Subsequent analysis with other multi- and single-locus systems, however, failed to confirm this preliminary result. The paternity probability computed on the basis of the single-locus systems was 0.99997, providing compelling evidence in favour of true paternity. The present case thus demonstrates that even when two mutations turn up in a DNA fingerprint, these may be readily recognized as such.

1993 ◽  
Vol 50 (7) ◽  
pp. 1559-1567 ◽  
Author(s):  
T. A. Stevens ◽  
R. E. Withler ◽  
S. H. Goh ◽  
T. D. Beacham

A multilocus DNA probe, B2-2, isolated from chinook salmon (Oncorhynchus tshawytscha) and a single-locus Atlantic salmon (Salmo salar) probe, 3.15.34, were examined for discriminatory ability among seven parents and 33–37 juveniles from five families of chinook salmon. DNA fingerprint patterns were observed in Hae III-digested chinook salmon DNA probed with B2-2. Between 8 and 20 fragments, from 2.20 kilobase pairs (kbp) to 19.0 kbp, were detected in each individual. The level of band sharing among unrelated parents was 0.18. Probe 3.15.34 hybridized with a total of nine DNA fragments, from 3.35 to 6.00 kbp, in the chinook salmon parents and progeny. One or two fragments were detected in each individual. Pedigree analysis confirmed that 3.15.34 detected both alleles of a single polymorphic locus whereas B2-2 detected autosomal, unlinked, predominantly heterozygous DNA fragments that were inherited in a Mendelian fashion at a minimum of 10 polymorphic loci. Among juvenile chinook salmon, levels of band sharing detected with probe B2-2 increased with increasing relatedness, and clustering based on differences in banding patterns distinguished unrelated progeny, half sibs, and full sibs even in the absence of parental genotypic data.


2011 ◽  
Vol 14 (3) ◽  
pp. 347 ◽  
Author(s):  
Natalie Rae Parfitt ◽  
Michael Frederick Skinner ◽  
Charles Bon ◽  
Isadore Kanfer

Purpose: Investigations were carried out to assess the use of tape stripping (TS) for the determination of bioequivalence of topical products containing 1% clotrimazole. Methods: The study design involved the establishment of an appropriate application time, which was determined by conducting a dose duration study. Subsequently, two bioequivalence studies were conducted: i) using the brand (Canesten Topical - 1% clotrimazole cream) as both the test and the reference product and ii) comparing Canesten cream with a gel product containing the same concentration of clotrimazole (1%). Each tape strip was individually analyzed for clotrimazole content using an HPLC method and Transepidermal Water Loss (TEWL) measurements were used to normalize the stratum corneum thicknesses between subjects. Results: The results of the TS investigations showed that, if the study is sufficiently powered, tape stripping may be used to determine bioequivalence according to the conventional bioequivalence limits of 0.8–1.25, as well as detect formulation differences between different clotrimazole products. Conclusions: The data from this study provided compelling evidence that tape stripping has the necessary attributes and potential to be used as a tool for the bioequivalence assessment of topical clotrimazole and/or other topical formulations, thereby circumventing the need to undertake expensive and time-consuming clinical trials for such products. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.


1998 ◽  
Vol 32 (1) ◽  
pp. 80-85 ◽  
Author(s):  
F. Benavides ◽  
D. Cazalla ◽  
C. Pereira ◽  
A. Fontanals ◽  
M. Salaverri ◽  
...  

A genetic monitoring of the BALB/c mouse foundation colony in our animal facility was carried out. The techniques of choice were skin grafting, coat colour test, flow cytometric analysis for H2 antigens (loci H2-D and H2-A), electrophoretic analysis of isoenzymes (loci Idh1, Pep3, Es3 and Mod1), PCR-amplified microsatellites (loci Igh-V, Ngfg, Plau, Crp, Igh, D16Mit5, D3Mit49 and D17Mit16) and DNA fingerprinting (multilocus probes 33.6, 33.15 and (CAC)5). No evidence of genetic contamination was found, ruling out the possibility of an outcross with AKR, the other albino strain maintained at the facility. Nevertheless, DNA fingerprint patterns revealed evidence of genetic heterogeneity in four out of nine lines of the nucleus colony, interpreted as minisatellite mutations favoured for a single line system with more than 40 generations of separation from the ancestral pair. These mice are mainly used in cancer and immunological research within the institute.


2016 ◽  
Vol 6 (3) ◽  
Author(s):  
Akiko Takashima ◽  
Naoki Takeshita ◽  
Toshihiko Kinoshita

A 41-year old pregnant woman underwent amniocentesis to conduct a conventional karyotyping analysis; the analysis reported an abnormal karyotype: 46,XY,add(9)(p24). Chromosomal microarray analysis (CMA) is utilized in prenatal diagnoses. A single nucleotide polymorphism microarray revealed a male fetus with balanced chromosomal translocations on 9p and balanced chromosomal rearrangements, but another chromosomal abnormality was detected. The fetus had microduplication. The child was born as a phenotypically normal male. CMA is a simple and informative procedure for prenatal genetic diagnosis. CMA is the detection of chromosomal variants of unknown clinical significance; therefore, genetic counseling is important during prenatal genetic testing.


1998 ◽  
Vol 88 (12) ◽  
pp. 1283-1293 ◽  
Author(s):  
S. Bentley ◽  
K. G. Pegg ◽  
N. Y. Moore ◽  
R. D. Davis ◽  
I. W. Buddenhagen

Genetic variation within a worldwide collection of 208 isolates of Fu-sarium oxysporum f. sp. cubense, representing physiological races 1, 2, 3, and 4 and the 20 reported vegetative compatibility groups (VCGs), was analyzed using modified DNA amplification fingerprinting. Also characterized were 133 isolates that did not belong to any of the reported VCGs of F. oxysporum f. sp. cubense including race 3 isolates from a Heliconia species and isolates from a symptomatic wild banana species growing in the jungle in peninsular Malaysia. The DNA fingerprint patterns were generally VCG specific, irrespective of geographic or host origin. A total of 33 different genotypes were identified within F. oxysporum f. sp. cu-bense; 19 genotypes were distinguished among the isolates that belonged to the 20 reported VCGs, and 14 new genotypes were identified among the isolates that did not belong to any of the existing VCGs. DNA fingerprinting analysis also allowed differentiation of nine clonal lineages within F. oxysporum f. sp. cubense. Five of these lineages each contained numerous closely related VCGs and genotypes, and the remaining four lineages each contained a single genotype. The genetic diversity and geographic distribution of several of these lineages of F. oxysporum f. sp. cubense suggests that they have coevolved with edible bananas and their wild diploid progenitors in Asia. DNA fingerprinting analysis of isolates from the wild pathosystem provides further evidence for the coevolution hypothesis. The genetic isolation and limited geographic distribution of four of the lineages of F. oxysporum f. sp. cubense suggests that the pathogen has also arisen independently, both within and outside of the center of origin of the host.


2002 ◽  
Vol 92 (7) ◽  
pp. 785-793 ◽  
Author(s):  
D. V. Phillips ◽  
I. Carbone ◽  
S. E. Gold ◽  
L. M. Kohn

Both typical late season stem infections and atypical early season rosette infections of canola, a relatively new crop in the southeastern United States, were caused by Sclerotinia sclerotiorum. The 51 DNA fingerprints (from 71 isolates) did not match any fingerprints from previous studies of canola or other crops. Single locus haplotypes from nuclear DNA sequences included 18 in the intergenic spacer (IGS) of the rRNA repeat, four in 44.11, six in translation elongation factor 1α, three in calmodulin (CAL), and two in chitin synthase 1. Contingency permutation testing for associations of infection type with DNA fingerprint, single- or multilocus haplotype, or hierarchically nested clades based on single locus haplotypes found significant association of haplotype with mycelial compatibility group and DNA fingerprint for all loci except CAL. Significant association of IGS haplotypes with symptom type was detected in one pathogen population. Southeastern U.S. canola was infected by both recently evolved, geographically dispersed pathogen genotypes and older, indigenous genotypes (Carbone and Kohn, 2001. Mol. Ecol. 10:947–964). Indigenous haplotypes are infection-type generalists, and the most frequently isolated from rosette infections. In contrast, haplotypes from the most recently evolved, dispersed population were associated one-to-one with infection type, with only the most recently evolved haplotypes infecting rosettes.


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