IR@PKUHSC  > 北京大学第一临床医学院  > 儿科
学科主题临床医学
Antigen retrieval with protease digestion applied in immunohistochemical diagnosis of Alport syndrome
Guan, Na; Yu, Li-Xia; Wu, Guo-Hong; Xing, Yan; Ding, Jie
关键词Alport syndrome antigen retrieval diagnosis paraffin-embedded renal sections protease digestion
刊名NEPHROLOGY DIALYSIS TRANSPLANTATION
2008-11-01
DOI10.1093/ndt/gfn305
23期:11页:3509-3513
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Transplantation ; Urology & Nephrology
资助者National Scientific Foundation of Chinese ; International Collaborative Genetic Research Training Grant ; NIH/FIC ; International Collaborative Genetics Research Training Program ; Pediatric Nephrology Department of Peking University First Hospital ; National Scientific Foundation of Chinese ; International Collaborative Genetic Research Training Grant ; NIH/FIC ; International Collaborative Genetics Research Training Program ; Pediatric Nephrology Department of Peking University First Hospital
研究领域[WOS]Transplantation ; Urology & Nephrology
关键词[WOS]RENAL BIOPSIES ; IV COLLAGEN ; IMMUNOFLUORESCENCE
英文摘要

Background. Immunofluorescence (IF) staining of type IV collagen alpha chains on fresh frozen renal tissue is a convenient and accurate diagnosis technique for Alport syndrome (AS), which is restricted in the application with a non-frozen section. An alternative method for a paraffin-embedded section is needed in order to extend the application in various specimens. In this study, immunohistochemical staining of type IV collagen alpha chains on paraffin-embedded renal sections was investigated.

Methods. Three antigen retrieval methods including autoclave heating, pepsin digestion and protease digestion were tried on paraffin-embedded renal sections from two X-linked male AS patients, two X-linked female AS patients and two autosomal recessive AS patients. Two patients with isolated haematuria were also involved. Normal portions of nephrectomized kidneys from two patients with renal tumours were used as controls. The immunohistochemical staining pattern of type IV collagen was compared with that of IF staining.

Results. The results showed that both the autoclave heating method and protease digestion can be used for antigen retrieval for type IV collagen alpha chains. Compared to autoclave heating, protease digestion had the advantage of being less time consuming, with less renal sections being lost from the slides; it was the most optimal antigen retrieval method in this study. After protease digestion for 40 min, type IV collagen alpha 3 and alpha 5 chains showed a clear and proper distribution pattern in AS patients, haematuria patients and controls, which is the same as that of IF staining.

Conclusions. Compared to the autoclave heating method, protease antigen retrieval is more convenient and effective and can be used to restore type IV collagen alpha chains on paraffin-embedded renal sections for the diagnosis of AS.

语种英语
所属项目编号39770780 ; 39970775 ; 30371495 ; 30400482 ; D43 TW06176
资助者National Scientific Foundation of Chinese ; International Collaborative Genetic Research Training Grant ; NIH/FIC ; International Collaborative Genetics Research Training Program ; Pediatric Nephrology Department of Peking University First Hospital ; National Scientific Foundation of Chinese ; International Collaborative Genetic Research Training Grant ; NIH/FIC ; International Collaborative Genetics Research Training Program ; Pediatric Nephrology Department of Peking University First Hospital
WOS记录号WOS:000260153600023
Citation statistics
Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/67845
Collection北京大学第一临床医学院_儿科
作者单位Peking Univ, Hosp 1, Dept Pediat, Beijing 100034, Peoples R China
Recommended Citation
GB/T 7714
Guan, Na,Yu, Li-Xia,Wu, Guo-Hong,et al. Antigen retrieval with protease digestion applied in immunohistochemical diagnosis of Alport syndrome[J]. NEPHROLOGY DIALYSIS TRANSPLANTATION,2008,23(11):3509-3513.
APA Guan, Na,Yu, Li-Xia,Wu, Guo-Hong,Xing, Yan,&Ding, Jie.(2008).Antigen retrieval with protease digestion applied in immunohistochemical diagnosis of Alport syndrome.NEPHROLOGY DIALYSIS TRANSPLANTATION,23(11),3509-3513.
MLA Guan, Na,et al."Antigen retrieval with protease digestion applied in immunohistochemical diagnosis of Alport syndrome".NEPHROLOGY DIALYSIS TRANSPLANTATION 23.11(2008):3509-3513.
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