Feliciani C, Di Muzio M, Mohammad Pour S, Allegretti T, Amerio P, Toto P, Coscione G, Proietto G, Amerio P
Department of Dermatology, University G.D'Ann unzio, Chieti, Italy.
J Eur Acad Dermatol Venereol. 1998 May;10(3):243-7.
Epidermolysis bullosa acquisita (EBA) and bullous pemphigoid (BP) are diseases with similar clinical, histological, and immunofluorescent findings. Diagnosis requires the use of immunoelectron microscopy, immunoprecipitation or immunoblotting, but in recent years the differential diagnosis has been based on a cheaper technique named salt split skin. This study demonstrates that with a suction blister the fracture is at the same level as that obtained with the sodium split method and that it is also faster and cheaper. Suction blisters on normal skin and autoimmune perilesional bullous lesions, obtained with a hand vacuum pump, were studied by direct immunofluorescence and electron microscopy to evaluate the level of the split on normal suction split skin. Normal human split skin was also used as a substrate for an indirect immunofluorescent study using sera of patients with BP (68 sera), EBA (10 sera) and cicatricial pemphigoid (CP) (16 sera). Direct immunofluorescent examination was also done on perilesional skin after artificial separation obtained with a hand-vacuum pump in patients with the same diseases listed above (32 BP, 11 CP, 6 EBA).
On normal human skin split by suction or sodium chloride (NaCl; 1 mol/l) direct immunofluorescence and electron microscopy demonstrated that the split is at the lamina lucida level. Indirect immunofluorescent study of both normal human skin and perilesional skin split using suction as a substrate showed IgG deposits localized on the floor of the suction blister in all cases of EBA, whereas in over 88% of cases of BP and in over 62% of CP the IgG were localized on the roof. Similar results were obtained with direct immunofluorescence in perilesional skin.
'Suction split' represents a simple technique to differentiate EBA from BP. This method provides final response in a few hours compared to at least 1-2 days with the sodium split method. Furthermore, the suction split method is cheaper and the tissue can be re-utilized for molecular biology and immunohistochemical studies.
获得性大疱性表皮松解症(EBA)和大疱性类天疱疮(BP)是临床、组织学及免疫荧光表现相似的疾病。诊断需要采用免疫电子显微镜、免疫沉淀或免疫印迹法,但近年来鉴别诊断基于一种名为盐裂皮肤的成本较低的技术。本研究表明,通过抽吸水疱获得的裂隙与用氯化钠分裂法获得的裂隙处于同一水平,而且速度更快、成本更低。用手动真空泵在正常皮肤和自身免疫性损害周围的大疱性皮损上制造抽吸水疱,通过直接免疫荧光和电子显微镜检查来评估正常抽吸分裂皮肤的分裂水平。正常人分裂皮肤还用作间接免疫荧光研究的底物,使用BP患者(68份血清)、EBA患者(10份血清)和瘢痕性类天疱疮(CP)患者(16份血清)的血清。对上述相同疾病患者(32例BP、11例CP、6例EBA)用手动真空泵进行人工分离后,对损害周围皮肤也进行了直接免疫荧光检查。
在通过抽吸或氯化钠(1mol/L)分裂的正常人皮肤上,直接免疫荧光和电子显微镜检查表明分裂位于透明层水平。以抽吸分裂的正常人和损害周围皮肤为底物进行的间接免疫荧光研究显示,在所有EBA病例中,IgG沉积物位于抽吸水疱的底部,而在超过88%的BP病例和超过62%的CP病例中,IgG位于顶部。在损害周围皮肤的直接免疫荧光检查中也得到了类似结果。
“抽吸分裂”是一种区分EBA和BP的简单技术。与氯化钠分裂法至少需要1 - 2天相比,该方法在数小时内即可得出最终结果。此外,抽吸分裂法成本更低,组织可重新用于分子生物学和免疫组织化学研究。