建立了一种应用近红外荧光免疫层析技术快速检测食源性甲型肝炎(甲肝)病毒的方法。采用近红外荧光染料标记甲肝病毒单克隆抗体,将甲肝病毒多克隆抗体和羊抗鼠IgG多克隆抗体喷涂于硝酸纤维膜上分别作为检测线(T线)和质量控制线(C线),研制检测甲肝病毒的近红外免疫层析试纸条及配套标准物质。研究表明,所建立的近红外免疫法对检测甲肝病毒具有良好的特异性和较高的灵敏度,最低检测限为1 μg/mL,与肠道病毒71型(EV71)、腺病毒、轮状病毒、诺如病毒无交叉反应。效果评价试验发现,与传统检测方法和胶体金法相比较,所建立的近红外荧光法检测时间最短,检测限低于胶体金法。近红外荧光免疫层析技术大大缩短了检测时间,可用于食品中甲肝病毒的高效检测,为食品安全监管提供了技术支持。
万晓楠
,
畅晓晖
,
齐玮
,
高欣
,
乔彬
,
杨向莹
,
李小林
,
张惠媛
,
石嵩
,
张捷
,
周熙成
. 基于近红外免疫层析技术快速检测食源性甲型肝炎病毒[J]. 食品与发酵工业, 2020
, 46(7)
: 213
-217
.
DOI: 10.13995/j.cnki.11-1802/ts.021114
A method for rapidly detecting hepatitis A virus by using near-infrared fluorescent immunoassay was established. A monoclonal antibody against hepatitis A virus was labeled with a near-infrared fluorescent dye. Polyclonal antibody and goat anti-mouse IgG polyclonal antibody were sprayed on a nitrocellulose membrane . Then those were used as test line and control line to develop NIR immunoassay strips for detecting hepatitis A virus. Studies have shown that NIR immunoassay has good specificity and high sensitivity for detecting hepatitis A virus. The minimum detection limit is 1 μg/mL and no cross reaction with EV71, adenovirus, rotavirus and norovirus. Through the effect evaluation test, it was found that NIR immunoassay method has the shortest detection time compared with the traditional and colloidal gold method. The detection limit is lower than the colloidal gold. Thus, NIR immunoassay can be used for detecting hepatitis A virus and provide reliable technical support for food safety supervision.
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