Thomas A S, Aaskov J G
Forensic Biology Section, State Health Laboratory, Brisbane, Queensland, Australia.
Forensic Sci Int. 1990 Sep;47(2):173-80. doi: 10.1016/0379-0738(90)90211-g.
The application of a polyacrylamide gel isoelectric focusing (PAGIEF) and immunoblotting procedure for the identification of native alpha 2HS-glycoprotein (AHSG) in routine casework blood stains has produced reportable results on 57.2% of samples. This reporting rate is lower than that for group specific component (GC) (83.8%) and phosphoglucomutase (PGM 1) (72.8%) phenotyping of the same samples. Blood stain samples were desialyzed with 1 U/ml neuraminidase, overnight at room temperature prior to PAGIEF in gels containing pharmalyte pH 5-6 and 2.5 M urea. Simple AHSG patterns were developed by immunoblotting. This procedure was five times as sensitive as the native AHSG method and desialyzation was reproducible over a range of incubation times and neuraminidase concentrations. The application of the desialyzed AHSG analysis to routine casework samples has resulted in a significant increase in the number of reportable results (762 reported out of 1027 samples). This reporting rate (74.2%) compares favourably with that for GC (79.1%) and PGH 1 (69.6%) phenotyping of the same samples. The three AHSG alleles (AHSG*1, 2 and 3) are clearly resolved after sample desialyzation and separation in gels containing pharmalyte pH 5-6 and 2.5 M urea. The sensitivity of desialyzed AHSG phenotyping approaches that of GC and this technique is worthy of inclusion in blood stain screening protocols of forensic laboratories in regions where the population has a limited range of rare AHSG alleles.
在常规案件工作血痕中，应用聚丙烯酰胺凝胶等电聚焦（PAGIEF）和免疫印迹程序鉴定天然α2HS-糖蛋白（AHSG），在57.2%的样本中产生了可报告结果。该报告率低于同一样本的群体特异性成分（GC）（83.8%）和磷酸葡萄糖变位酶（PGM 1）（72.8%）表型分析的报告率。在含有pH 5 - 6 Pharmalyte和2.5 M尿素的凝胶中进行PAGIEF之前，将血痕样本用1 U/ml神经氨酸酶在室温下脱唾液酸过夜。通过免疫印迹产生简单的AHSG图谱。该程序的灵敏度是天然AHSG方法的五倍，并且在一系列孵育时间和神经氨酸酶浓度范围内，脱唾液酸是可重复的。将脱唾液酸AHSG分析应用于常规案件工作样本，使可报告结果的数量显著增加（1027个样本中有762个报告结果）。该报告率（74.2%）与同一样本的GC（79.1%）和PGH 1（69.6%）表型分析的报告率相比具有优势。在含有pH 5 - 6 Pharmalyte和2.5 M尿素的凝胶中对样本进行脱唾液酸和分离后，三个AHSG等位基因（AHSG*1、2和3）清晰可辨。脱唾液酸AHSG表型分析的灵敏度接近GC，在人群中罕见AHSG等位基因范围有限的地区，该技术值得纳入法医实验室的血痕筛查方案。
