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PubMed2004

Molecular medicine: a primer for clinicians. Part XIV: Stem cells.

Earlier papers in this series have described many of the tools of molecular biology that are finding practical bedside applications. We have reviewed how molecular diagnostics, genetic testing, and DNA and protein microarrays all have become part of clinical medicine. We have also described the potential of the human genome project to impact the practice of medicine. Finally, we have discussed several ethical issues, including genetic screening, forensic DNA testing, and gene therapy. In this paper, and the next, we will discuss two topical areas in which molecular biology may influence how medicine is practiced in the future; stem cell research and cloning technology.

Embryo, Mammalian↗
PubMed2006

[Developing of a new multicolor-fluorescent labeled STR amplification kit].

OBJECTIVE: To develop a PCR-based STR system for genotyping of 18 loci (Amelogenin, D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D16S539, TH01, TPOX, CSF1PO, D7S820, D2S1338, D19S433, D12S391 and D19S253). METHODS: By using primers labeled with four color fluorescent (FAM, HEX, TAMRA and ROX), two multiplex amplification reaction systems were developed to genotype Amelogenin and 17 STR loci. RESULTS: Amelogenin and these 17 STR loci were genotyped successfully in different kinds of biological samples by the kit. CONCLUSION: The STR amplification kit developed in our study gives a new approach to genotype these 18 loci in a efficient, steady and reliable way.

Alleles↗
PubMed2006

[The application of PCR-SSCP in forensic mtDNA typing].

OBJECTIVE: To study the application of PCR-SSCP in forensic mtDNA typing. METHODS: Primers flanking the mtDNA HV-I and HV-II regions were designed. By PCR-SSCP techniques, 70 family trios and 140 unrelated Wuhan Han individuals were investigated and analyzed. RESULTS: In 70 family trios, the SSCP profiles in region HV-I and HV-II of children were not same to that of their fathers in 98.57% and 97.13% respectively but were identical with their mothers. In 140 unrelated Wuhan Han individuals, 21 haplotypes were found in HVI, GD = 0.9556; 16 haplotypes were found in HVII, GD = 0.9356. CONCLUSION: PCR-SSCP technique may be useful in forensic mtDNA typing, especially for screening the suspects.

DNA Fingerprinting↗
PubMed2006

[Recent advancement in miniSTR research].

As the most popular and important inherited marker used in forensic identification, short tandem repeat (STR) always have partial DNA profiling or even no results when handling degraded or minute DNA sample. Through redesigning primers close to STR core repeats, MiniSTR can access shorter STR loci and increase success rate of DNA profiling in degraded or minute DNA sample. The review provides an update on the advancement of miniSTR research to give information in the practice of forensic science.

DNA Degradation, Necrotic↗
PubMed2006

[Allele frequencies and species specificity of five short tandem repeat loci of Chinese Han population in Chengdu].

OBJECTIVE: To obtain the data in polymorphism distribution of the five short tandem repeat (STR) loci: D18S979, D11S2014, D18S548, D1S1667 and GATA164F07 of Chinese Han population in Chengdu, and to evaluate their usefulness in the field of species specificity in forensic science. METHODS: PCR, polyacrylamide gel electrophoresis (PAGE) and silver staining techniques were used to analyze the DNA samples from 100 unrelated individuals of Chinese Han ethnic group in Chengdu. Twelve different animals: monkey, pig, dog, bull, goat, chicken, duck, eel, mudfish, rabbit, guinea pig and mouse were selected as controls in this study for evaluating the species specificity of the five STR loci. RESULTS: Six alleles and twelve genotypes were observed in D18S979. Five alleles and eleven genotypes were observed in D11S2014. Five alleles and thirteen genotypes were observed in D18S548. Seven alleles and nineteen genotypes were observed in D1S1667. Six alleles and fourteen genotypes were observed in GATA164F07. The genotype distributions of the five loci were analyzed by some related software and no deviation from the Hardy-Weinberg equilibrium was observed. Evaluated by way of using different animals as controls, monkey had amplification products at the extra-typing field of D18S979, D11S2014 and D1S1667. Bull, dog and eel had amplification product at typing field of D18S979, and pig, duck, mouse and rabbit had weak product. Bull had weak product at the typing field of D18S548. Dog, goat and eel had product at the typing field of D1S1667. Dog had weak product at the typing field of GATA164F07. Mudfish, chicken and guinea pig had no amplification product at the five loci. CONCLUSION: These data indicate that D18S979, D18S548, D1S1667 and GATA164F07 are highly polymorphic and D11S2014, D18S548 and GATA164F07 can play a key role in species identification.

Animals↗
PubMed1989

Improved genomic blot hybridization.

For restriction fragment length polymorphism (RFLP) and other molecular biology experiments, well prepared (undamaged) genomic DNA is needed, which is then digested with restriction enzymes, transferred to nitrocellulose paper and hybridized with radioactive or nonradioactive probe. DNA damage occurs in fields such as clinical genetics and forensic medicine in the form, for example, of single stranded breaks. If these DNAs are treated with T4 DNA ligase prior to restriction endonuclease digestion, the damaged DNA is repaired. This "repaired DNA" gives improved Southern hybridization results with single copy gene or repeated genes.

Biotechnology↗
PubMed1976

Immunofixation. I. General principles and application to agarose gel electrophoresis.

Immunofixation offers the worker an economical means of physically locating a protein in an electrophoretic strip and is ideally suited to forensic medicine, genetic studies, or research. The method is as simple and economical as the commonly used one- or two-dimensional immunoelectrophoresis, yet yields considerably more information.

Blood Protein Electrophoresis↗
PubMed1995

The blood revolution initiated by the famous footnote of Karl Landsteiner's 1900 paper.

A 1900 publication authored by Karl Landsteiner, at the age of 32 years, contained a footnote which stated that, "the serum of healthy human beings not only agglutinates animal red cells, but also often those of human origin, from other individuals". He followed up this statement in his 1901 paper, and concluded that, "My observations reveal characteristic differences between blood serum and red blood cells of various apparently healthy persons" and that "the reported observations may assist in the explanation of various consequences of therapeutical blood transfusions". These significant observations resulted in the discovery of A, B, O and AB blood groups and later led to successful blood transfusions in humans. The impact of the revolutionary finding by Landsteiner also changed a number of biomedical disciplines such as immunochemistry, medical anthropology, forensic medicine, genetics and pathology.

Allergy and Immunology↗
PubMed2004

[Genetic polymorphism of 3 STR loci on chromosome X and its forensic application in Chinese population].

To investigate the genetic polymorphisms of three short tandem repeats loci of chromosome X in Chinese Han population in Chengdu area and its use in forensic science. Three X-chromosome linked short tandem repeat loci were analyzed by PCR followed by polyacrylamide gel electrophoresis. Hardy-Weinberg equilibrium was tested and forensic interested value was calculated . The power of exclution of DXS6804, DXS9896 and GATA144D04 is 0.5990, 0.6220, 0.4280, respectively. The result showed that all the three STR loci were polymorphic among 100 unrelated females and 120 unrelated males from Chinese Han population. Chi(2) tests demonstrated that genotype frequencies in females did not depart from Hardy-Weinberg equilibrium. Three X-chromosome linked short tandem repeat loci have high polymorphism, they can be applied to forensic medicine and population genetics.

Alleles↗
PubMed1995

Flow chart HLA-DQA1 genotyping and its application to a forensic case.

The detection of genetic polymorphism has become increasingly important in forensic science as well as in medical genetics. In this report, we describe a systematic flow chart system for HLA-DQA1 genotyping by an improved PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method coupled with the PRSM (PCR-mediated restriction site modification) method. This flow chart typing system can easily discriminate between a total of eight reported DQA1 alleles commonly found in Chinese. We have applied this flow chart typing system in a forensic case as well as in the determination of the frequencies of the eight DQA1 alleles in 121 unrelated Taiwan Chinese subjects. Our results show that the flow chart DQA1 genotyping is a simple, fast, and accurate system which, in the future, may be considered as an alternative method for routine individual identification in forensic casework, and for paternity testing and tissue typing in medical genetics.

Alleles↗
PubMed1990

Amplification of a variable number of tandem repeats (VNTR) locus (pMCT118) by the polymerase chain reaction (PCR) and its application to forensic science.

A genetic locus (D1S58, defined by DNA probe pMCT118) that contains a variable number of tandem repeats (VNTR) has been successfully amplified from a very small amount of genomic deoxyribonucleic acid (DNA) by the polymerase chain reaction (PCR). The DNA sequence of the locus was determined and was found to consist of a 16-base consensus sequence and flanking sequences. Oligonucleotide primers complementary to the flanking sequences were synthesized to serve as primers for amplification of MCT118 by the PCR method. Human genomic DNA isolated from blood (2 ng from each sample) was successfully amplified at the MCT118 locus, and polymorphic bands were detectable by ethidium bromide staining after electrophoresis on polyacrylamide gels. Determination of genotypes at this VNTR locus can now be routinely achieved within 24 h, without the need for Southern blots or radioactive materials. Furthermore, the small size (387 to 723 base pairs) of the DNA fragments produced in the PCR amplification permits good resolution of individual alleles that differ by only one repeat unit. The precise specification of the number of tandem repeats present in each allelic fragment is reproducible from one analysis to another.

Alleles↗
PubMed2005

Population genetic study in two Transylvanian populations using forensically informative autosomal and Y-chromosomal STR markers.

Our study provides population genetic data on two population samples collected in a Hungarian speaking region of Transylvania, Romania. Allele frequency and profile databases were generated on 17 autosomal STR loci (D2S1338, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D19S433, D21S11, VWA, FGA, TH01, TPOX, CSF1PO, Penta E and Penta D) as well as at the 12 European Y-STR extended haplotype loci (DYS19, DYS389-I/II, DYS390, DYS391, DYS392, DYS393, DYS385 loci, DYS437, DYS438 and DYS439). Data were compared to a Central Hungarian (Budapest region) population sample [B. Egyed, S. Füredi, M. Angyal, L. Boutrand, A. Vandenberghe, J. Woller, Z. Padar, Analysis of eight STR loci in two Hungarian populations, Forensic Sci. Int. 113 (2000) 25-27] that was used as a reference group of the Hungarian population. Calculating the F(ST) indices and with the pairwise comparisons of interpopulation molecular variance (AMOVA) the two populations from Transylvania could be fit into the Hungarian population data showing less substructuring effects as compared to the previous findings in Hungary [B. Egyed, S. Füredi, M. Angyal, L. Boutrand, A. Vandenberghe, J. Woller, Z. Padar, Analysis of eight STR loci in two Hungarian populations, Forensic Sci. Int. 113 (2000) 25-27; B. Egyed, S. Füredi, M. Angyal, I. Balogh, L. Kalmar, Z. Padar, Analysis of the population heterogeneity in Hungary using fifteen forensically informative STR markers, Forensic Sci. Int. 158 (2005) 244-249].

Chromosomes, Human, Y↗
PubMed2004

[Genetic polymorphisms of three STR loci on chromosome X and their forensic application in a Chinese Han population].

OBJECTIVE: To understand the allele structure and genetic polymorphism at three STR loci on chromosome X in Chinese Han population and make an evalution of their forensic application. METHODS: EDTA-blood samples were collected from the unrelated individuals in Chengdu, China. After being extracted with Chelex method, the DNA samples were amplified by PCR technique. The PCR products were analyzed by PAG electrophoresis and the approach of automated fluorescence detection. Hardy-Weinberg equilibrium of females was tested and every forensic interested value was calculated. RESULTS: The polymorphisms of all 3 STR loci were obtained from 100 unrelated females and 120 unrelated males from Chinese Han ethnic group. Chi-square tests on the genotype frequencies in females did not reveal deviations from Hardy-Weinberg equilibrium. CONCLUSION: The obtained data are beneficial to understanding the population genetics of the three STR loci in Chinese Han population. For forensic genetics, the obtained data can be used to calculate the probabilities dealing with the paternity test and the individual identification.

China↗