贝类中3种组织血型抗原ELISA检测方法的建立与分型

Use of ELISA for the detection and typing of three HBGAs in shellfish

人类组织血型抗原(histo-blood group antigens,HBGAs)是诺如病毒(NoVs)的结合受体,本研究假设贝类中也存在类似的HBGAs并且特异性地富集NoVs,利用HBGAs单克隆抗体,建立贝类中3种HBGAs的ELISA检测方法,分析牡蛎(Crassostrea gigas)、缢蛏(Sinonovacula constricta)、菲律宾蛤仔(Ruditapes philippinarum)、紫贻贝(Mytilus galloprovincialis)、毛蚶(Scapharca subcrenata)、栉孔扇贝(Chlamys farreri)等6种双壳贝类中HBGAs的类型。结果显示,在上述6种贝类中都检测出A型抗原,其中菲律宾蛤仔的检出率为11.6%(9/77),紫贻贝的检出率为28.1%(16/57),缢蛏样品为72.3%(47/65),栉孔扇贝为84.6%(58/69),其余贝类检出率为100%;只有牡蛎样品检出H抗原,检出率为30.7%(28/91);在缢蛏和毛蚶样品中检测出B型抗原,检出率分别为76.9%(50/65)和77.8(56/72)。结果表明贝类中存在不止1种类型的HBGAs。

The histo-blood group antigens（HBGAs） located on cells of the human gastrointestinal tract function as receptors for NoVs.It is hypothesized that HBGA-like receptors are present in shellfish and are responsible for the specific accumulation of NoVs.We used human HBGA-specific monoclonal antibodies（MAbs） to develop an ELISA method for detection of three forms of HBGA in six bivalve shellfish species（oysters,razor clam,clam, blue mussel,blood clam,and scallop）.All six species expressed type-A HBGAs.The detection rate of type-A HBGAs was 11.6%,28.1%,72.3%,and 84.6% in Ruditapes philippinarum,Mytilus galloprovincialis, Sinonovacula constricta,and Chlamys farreri tissue,respectively.All samples of oyster and blood clam contained type-A HBGAs.Type-H HBGAs were found only in oysters with a detection rate of 30.7%.Type-B HBGAs were present in the razor clam and blood clam（detection rate of 76.9% and 77.8%,respectively）.Our results suggest that multiple HBGAs are expressed in bivalve shellfish and provide insight into the mechanism of accumulation of NoVs by bivalve shellfish.