Genetic markers in human bone: II. Studies on ABO (and IGH) grouping.

A combination absorption-elution, two-dimensional absorption-inhibition procedure was used to determine the ABH antigen composition of a series of human bone specimens of known ABO type that had been aged up to nine months under dry and humid conditions at ambient temperature, 37 degrees C, and 56 degrees C; at ambient temperature in dry and wet soil; and buried in soil outdoors. Grouping data for the separate elution and inhibition testing, as well as for the combination procedure, are given. The combination method was found to be a highly reliable procedure for bone tissue ABH typing. Some data on microbial contaminants of human bone specimens aging in soil, and their effects on ABH typing results, are presented. No direct correlation between the properties of microbial contaminants and specific changes in the ABH antigenic composition of aging bone tissue specimens could be ascertained. Data on IGH antigen determination and on the quantitation of immunoglobulin G (IgG) in human bone tissue extracts indicated that immunoglobulin levels were typically too low to expect routinely successful Gm antigen testing results. However, these factors can sometimes be determined in fresh bone tissue extracts, particularly if the extracts are concentrated.

ABH antigen typing in bone tissue.

Results obtained from the ABH grouping of bone tissues by the absorption-elution procedure and by a recently described two-dimensional absorption-inhibition procedure are reported. Neither the elution nor the inhibition procedure alone yielded uniformly correct results. A combination procedure consisting of the use of both absorption-elution and two-dimensional absorption-inhibition is proposed for bone ABH grouping. When elution and inhibition were used in combination, specimens yielding concordant results with both techniques were correctly grouped.

Studies on ABH antigen grouping of ammoniacal extracts of bloodstains by absorption-elution.

The effects of absorption time, titer of antiserum or lectin in the absorption stage, and elution temperature on relative antibody or lectin yield in elutes in the absorption-elution procedure were studied in bloodstains and ammoniacal extracts of bloodstains using conventional grouping reagents. In addition, monoclonal anti-A and anti-B and affinity-purified Ulex europaeus agglutinin I (UEA I) reagents were employed for comparison in these processes. Ammoniacal extracts of bloodstains and dried bloodstains on cotton substrata behaved comparably with respect to the parameters studied. The monoclonal anti-A and anti-B and UEA I reagents studied yielded satisfactory results, comparable in some cases to conventional reagents, with respect to the parameters studied.

Simultaneous identification and determination of species origin, ABH antigens and isoenzyme markers in the same bloodstain.

A reliable procedure for the simultaneous identification of blood, determination of species origin and ABH antigens, and typing of isoenzyme markers from a sample of three 1-cm threads is described. Results obtained from known control as well as from casework bloodstains using this procedure were consistent with those obtained in parallel, conventional, individual tests under blind trial conditions.

Evaluation of antisera for bloodstain grouping. II. Ss, Kell, Duffy, Kidd, and Gm/Km.

Thirty-one different examples of commercially available blood grouping antisera specific for the S, s, K, k, Fya, Fyb, Jka, and Jkb antigens and anti-human globulin sera were serologically evaluated with red cells and in absorption-elution tests to determine their applicability to bloodstain antigen determinations. Nineteen examples of commercially available antisera specific for various Gm and Km antigens and their corresponding anti-D reagents were likewise evaluated in inhibition tests with sera and bloodstains. Elution tests with the blood grouping antisera and inhibition tests with the Gm/Km antisera on a series of aging bloodstains on cotton cloth, and on bloodstains on a number of different substrata, demonstrated that properly evaluated commercial antisera are useful reagents for bloodstain grouping in forensic serology.