Immunological detection of human phosphoglucomutase (PGM 1) subtypes

Anti-phosphoglucomutase (PGM) antibodies have been produced by immunising a sheep with a purified preparation of rabbit skeletal muscle PGM and used to devise an immunological procedure for detecting PGM isozymes after isoelectric focusing. The anti-rabbit PGM antibodies cross react with human PGM and can be used to identify the PGM1 isozymes characteristic of this polymorphism. The patterns revealed by immunodetection are exactly comparable with those obtained by isozyme staining.     

Identification of a phosphoglucomutase 1 (PGM 1) variant in a case of murder/suicide

A murder/suicide case is reported in which a phosphoglucomutase (PGM) 1*W9 variant was detected in a woman, her child, and from blood collected at the scene.     

Simultaneous identification of alpha-L-fucosidase and phosphoglucomutase (PGM) subtyping in semen stains

Seminal fluid and stains were analyzed by isoelectric focusing to determine the donor phenotype in the alpha-L-fucosidase (AlFuc) polymorphic system. The enzyme is found in both seminal fluid and spermatazoa. Three common phenotypes exist and can be identified in fluid specimens stored at 4 degrees C for more than a year. Untreated semen specimens display more than eight distinct bands of alpha-L-fucosidase activity with isoelectric points of pH 6.6 and below. Neuraminidase-treated specimens have enhanced banding patterns cathodally with a loss of activity in anodal bands making it easier to phenotype specimens. Semen stains maintained in dehumidified chambers at 25 or 37 degrees C retained activity for at least one month and could be accurately phenotyped. Activity was observed in semen specimens maintained at -20 degrees C in the dried state for a period of one year, whereas a complete loss of activity was observed after two weeks in similar specimens maintained at 25 or 37 degrees C under humid conditions. Of seventy-four semen stains analyzed, two had no apparent activity. Of the remaining seventy-two specimens 56, 32, and 12% were phenotyped as FUC 1-1, FUC 2-1, and FUC 2-2, respectively. Calculated gene frequencies are FUC1 = 0.72 and FUC2 = 0.28. Following analysis of alpha-L-fucosidase, the agarose gel can be chemically developed to reveal the PGM1 subtyping pattern. The ability to phenotype both systems in semen stains significantly improves the ability of the analyst to individualize this type of physical evidence. The probability of discrimination for these two combined systems is approximately 0.89.     

PGM1 subtypes determined by agarose gel isoelectrofocusing

PGM1 subtypes were determined in red cell hemolysates by isoelectric focusing on agarose gel plates. By this modified procedure PGM1 subtypes may be readily classified. Nine of the 10 expected phenotypes were found in a sample of 470 unrelated individuals from Southern Germany. The frequencies for the four alleles were found to be: PGM1(1+) = 0.212, PGM1(1-) = 0.1224, PGM1(2+) = 0.2043, PGM1(2-) = 0.0521.     

Rare PGM1 (6) and PGM1(7) alleles in the Polish population

Rare PGM1 phenotypes, 6-1, 6-2 and 7-2, were detected in blood samples from 3,437 non-related adults using electrophoresis in starch-gel and cellulose acetate membranes. Frequencies of 0.0009 and 0.0008 were calculated for PGM1(6) and PGM1(7), respectively, for a population from northern Poland. The variants had been inherited, which was confirmed by family studies.     

Determination of phosphoglucomutase (PGM1), acid phosphatase (ACP), and esterase D (ESD) in human bloodstains by hybrid isoelectric focusing (HIEF)

PGM1, ESD, and ACP were determined in bloodstain extracts by isoelectric focusing (IEF) with carrier ampholytes (CA) and HIEF. HIEF yields superior results in PGM typing from bloodstain extracts, whereas for ESD and ACP typing isoelectric focusing with carrier ampholytes seems to be the method of choice.     

Isoelectric focusing studies on the PGM1 subtypes in the northern Japanese population

The distribution of the human red cell phosphoglucomutase (PGM1) subtypes in samples from Japanese population (n = 277) living in the Miyagi Prefecture, the northern part of Japan, was investigated by applying the thinlayer polyacrylamide gel isoelectric focusing. In our population sample all the ten common phenotypes were demonstrated, and the estimated allele frequencies for the genes PGM1+1, PGM1-1, PGM2+1, and PGM2-1 were 0.671, 0.107, 0.161, and 0.061, respectively. Family studies (n = 40) indicated an autosomal codominant inheritance and confirmed the four alleles. The new system will increase the probability of exclusion in paternity cases among Japanese to 29.4% compared with 14.3% if the two allele system is used.     

Non-equilibrium pH gradient electrophoresis (NEPHGE) on ultrathin polyacrylamide gels containing separators: improved erythrocyte phosphoglucomutase (PGM) and esterase D (EsD) diagnosis in red cell lysates and bloodstains

Two rapid and reliable electrophoretic techniques for PGM1 and EsD typing on ultrathin polyacrylamide gels are described. They have been based on non-equilibrium pH gradient electrophoresis and on the addition of chemical spacers (EPPS for PGM1 and HEPES for EsD) to the gel mixture.     

Use of histoelectrofocusing in the determination of PGM1 subtypes in human body tissues

The present study reports the successful application of histo-electrofocusing to the determination of PGM1 subtypes in various tissues of the human body. The method described here is of practical use in individualizing parts of cadavers stored for up to 1 week.     

A comparison of the sensitivity of typing group-specific component (Gc) and the phosphoglucomutase (PGM1) locus in control and casework bloodstains by three isoelectric focusing methods

The performance of typing group-specific component (Gc) in bloodstains by two isoelectric focusing methods followed by its detection with silver staining has been compared with an established forensic system of typing phosphoglucomutase (PGM1) locus phenotypes by isoelectric focusing (IEF) in 1 mm gels. For Gc typing ultra-thin isoelectric focusing (UTIEF) gels and immobilized pH gradient (IPG) gels were used. Both laboratory prepared stains and casework stains were examined. The Gc UTIEF method is approximately eight times more sensitive than the existing PGM1 1 mm IEF method for control and casework stains. However, on average, a larger amount of stain was taken from casework stains than control stains for each typing system. A total of 53 casework stains were examined. Comparable success rates of 62% and 64% were obtained for typing Gc on UTIEF gels and PGM1 by 1 mm IEF, respectively. A success rate of 55% was obtained for typing Gc on IPGs. Bloodstains that were over 200 days old were successfully grouped by all three methods.     

Evaluation of a nonequilibrium isoelectric focusing (IEF) method for the simultaneous typing of esterase D (EsD), red cell acid phosphatase (AcP1), phosphoglucomutase (PGM1), adenylate kinase (AK), and adenosine deaminase (ADA)

A nonequilibrium isoelectric focusing method incorporating the chemical spacers MOPS and HEPES was developed and subsequently evaluated for its ability to reliably discriminate common and rare phenotypes in the esterase D (EsD), red cell acid phosphatase (AcP1), phosphoglucomutase (PGM1), adenylate kinase (AK), and adenosine deaminase (ADA) isoenzyme systems. The validation procedures used were blind testing, comparison of results to conventional methods, and evaluation of known rare variant phenotypes. This method proved to be a quick and reliable method for typing all five isoenzyme systems, while providing an excellent probability of discrimination (PD = 0.96).     

Detection of the rare PGM1(3) allele. Further biochemical and genetic characterization of the PGM1(3) isozymes

A family possessing the rare PGM1(3) allele has been found in North Carolina, and criteria for the electrophoretic separation and accurate typing of the PGM1(3) isozymes are outlined. The PGM1(3) isozymes detected proved to be useful in helping to determine parentage in an incest investigation. The pattern of segregation of the PGM1(3) allele in four generations of this family and thermostability studies on the PGM1(3) isozymes are presented.     

Alpha-1-antitrypsin (PiM) subtypes in Japanese

Using isoelectric focusing (IEF) in thin-layer polyacrylamide gel, the polymorphism of the serum alpha-1-antitrypsin (Pi system) was investigated in 335 healthy unrelated Japanese individuals living in Miyagi prefecture, the northern part of Japan. Six common and five rare variant phenotypes were identified in our population samples, and the estimated allele frequencies for the genes Pi^M1, Pi^M2 and Pi^M3 were 0.718, 0.238 and 0.044, respectively. Family studies (n = 46) showed an autosomal codominant inheritance, and no exclusion was found in 23 mother-child pairs.     

Diversity of 17-locus Y-STR haplotypes in Upper (Southern) Egyptians

Seventeen Y-STR loci included in the AmpF?STR^® Yfiler™ PCR Amplification kit were typed in a population sample of 208 males from Upper (South) Egypt. Of 204 observed haplotypes, 200 were unique (96.6%) and 4 were found twice each. The 17 loci gave a discriminating power of 0.9998. DYS458 showed the highest diversity as a single-locus marker (h = 0.868) along with a high frequency of microvariants and new alleles (22% of the sample). Other loci revealed duplicated and null alleles. Comparative analysis with Y-STR datasets of relevant populations and submission of the haplotypes to the Y-STR Haplotype Reference Database (YHRD) were undertaken.