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学科主题: 临床医学
题名:
Antigen retrieval with protease digestion applied in immunohistochemical diagnosis of Alport syndrome
作者: 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
发表日期: 2008-11-01
DOI: 10.1093/ndt/gfn305
卷: 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
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
项目资助者: 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
WOS记录号: WOS:000260153600023
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/67845
Appears in Collections:北京大学第一临床医学院_儿科_期刊论文

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作者单位: Peking Univ, Hosp 1, Dept Pediat, Beijing 100034, Peoples R China

Recommended Citation:
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.
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