水产学杂志
水產學雜誌
수산학잡지
CHINESE JOURNAL OF FISHERIES
2015年
2期
6-11
,共6页
哈维氏弧菌%胶体金%免疫层析%试纸条
哈維氏弧菌%膠體金%免疫層析%試紙條
합유씨호균%효체금%면역층석%시지조
Vibrio harveyi%colloidal gold%immunochromatography%test strip
用18~20nm 胶体金标记抗哈维氏弧菌 Vibrio harveyi 抗体并制备金标垫，分别将羊抗兔 IgG 和纯化后的抗哈维氏弧菌抗体包被于 NC 膜上作为质控线和检测线，组装制作了哈维氏弧菌胶体金免疫层析试纸条，优化了该试纸条的制作条件，测试了其性能。结果表明：所制备的哈维氏弧菌免疫层析试纸条具有较好的特异性，与费尼斯弧菌 Vibrio furnissii、维氏气单胞菌 Aeromonas veronii、美人鱼发光杆菌 Photobacterium damselae、类志贺邻单胞菌 Plesiomonas Shigelloides、鱼肠道弧菌 Vibrio ichthyoenteri、鳗利斯顿氏菌 Listonella anguillarum、迟缓爱德华氏菌 Edwardsiella tarda、海豚链球菌 Streptococcus iniae、嗜水气单胞菌 Aeromonas Hydrophila 等9种常见水产病原菌无明显交叉反应，检测灵敏度为3×105CFU/mL，5~10min 即可得出检测结果，具有快速、简便、特异性强和适应基层推广应用等优点，可用于养殖鱼类病原哈维氏弧菌的现场检测。
用18~20nm 膠體金標記抗哈維氏弧菌 Vibrio harveyi 抗體併製備金標墊，分彆將羊抗兔 IgG 和純化後的抗哈維氏弧菌抗體包被于 NC 膜上作為質控線和檢測線，組裝製作瞭哈維氏弧菌膠體金免疫層析試紙條，優化瞭該試紙條的製作條件，測試瞭其性能。結果錶明：所製備的哈維氏弧菌免疫層析試紙條具有較好的特異性，與費尼斯弧菌 Vibrio furnissii、維氏氣單胞菌 Aeromonas veronii、美人魚髮光桿菌 Photobacterium damselae、類誌賀鄰單胞菌 Plesiomonas Shigelloides、魚腸道弧菌 Vibrio ichthyoenteri、鰻利斯頓氏菌 Listonella anguillarum、遲緩愛德華氏菌 Edwardsiella tarda、海豚鏈毬菌 Streptococcus iniae、嗜水氣單胞菌 Aeromonas Hydrophila 等9種常見水產病原菌無明顯交扠反應，檢測靈敏度為3×105CFU/mL，5~10min 即可得齣檢測結果，具有快速、簡便、特異性彊和適應基層推廣應用等優點，可用于養殖魚類病原哈維氏弧菌的現場檢測。
용18~20nm 효체금표기항합유씨호균 Vibrio harveyi 항체병제비금표점，분별장양항토 IgG 화순화후적항합유씨호균항체포피우 NC 막상작위질공선화검측선，조장제작료합유씨호균효체금면역층석시지조，우화료해시지조적제작조건，측시료기성능。결과표명：소제비적합유씨호균면역층석시지조구유교호적특이성，여비니사호균 Vibrio furnissii、유씨기단포균 Aeromonas veronii、미인어발광간균 Photobacterium damselae、류지하린단포균 Plesiomonas Shigelloides、어장도호균 Vibrio ichthyoenteri、만리사돈씨균 Listonella anguillarum、지완애덕화씨균 Edwardsiella tarda、해돈련구균 Streptococcus iniae、기수기단포균 Aeromonas Hydrophila 등9충상견수산병원균무명현교차반응，검측령민도위3×105CFU/mL，5~10min 즉가득출검측결과，구유쾌속、간편、특이성강화괄응기층추엄응용등우점，가용우양식어류병원합유씨호균적현장검측。
A rapid, accurate and easy colloidal gold immunochromatographic test strip was developed for detection of Vibrio harveyi, in which the anti-V. harveyi polyclonal antibody coded A483 was conjugated with colloidal gold (18~20 nm) as signal generator on conjugate pad. The polyclonal antibody and goat anti-rabbit IgG were separately coated onto the same nitrocellulose membrane at the test line(T) for sample detection and the control line(C) for quality-control, respectively. The nitrocellulose membrane, gold conju-gate pad, sample pad, filter paper and absorbent pad were assembled to prepare the strips. The evaluation of detection specificity and sensitivity of this strip revealed that the test strips had the detection sensitivity of 3×105 CFU/mL for V. harveyi, no cross-reaction with other nine bacteria of aquaculture pathogens such as V. furnissii,Aeromonas veronii,Photobacterium damselae,Plesiomonas shigelloides, V. ichthyoenteri,Listonella anguillarum,Edwardsiella tarda,Streptococcus iniae and A. hydrophila. Accurate reading time for confirmation of the assay was completed in 5~10 min. The test strip generated in this study can be applied in on-spot detection of V. harveyi as a useful technique for rapid diagnosis of V. harveyi infection with the advantages of simple, rapid and without requirement of sophisticated equipment.
