UT-BioMED™

乙型流感病毒检测试剂盒（ELISA）说 明 书

【用 途】
本试剂盒采用双抗体夹心检测乙型流感病毒（Inf.B）抗原方法，操作过程采用一步法。在
微孔条上包被纯化的Inf.A单抗，检测时将待检病毒样本和酶标记抗体同时加入，温育洗板后加
入底物显色，终止后读值再判定结果。

【规 格】48人份

【试剂组成】
微孔包被板条（12×4条） 1块
酶标记物3.0ml 1瓶
阳性对照0.5ml 1支
阴性对照0.5ml 1支
浓 缩 洗涤 液 20 ml 1瓶
底物A 3. 0ml 1瓶
底物 B3. 0ml 1瓶
终止液3. 0ml 1瓶
封口膜 2个
自封袋 1个

【贮 藏】试剂盒置2～8℃保存；有效期为9个月。

【操作步骤】
1. 测前准备:实验前20分钟从冰箱中取出试剂盒，平衡至室温（18～25℃）；同时将浓缩洗涤液
用双蒸水1:25稀释后使用。
2. 用无菌棉拭子擦拭病人咽喉，采集后将棉拭子在装有0.5ml无菌生理盐水的试管中充分洗涤，
贴壁挤干，丢弃棉拭子。
3. 取咽拭子样本或阴/阳性对照各加50μl到反应孔内，再加入酶标记物50μl，震荡混匀，置37
℃温箱反应30分钟。
4. 洗板5次，扣干。
5. 将底物A、B各加50μl到反应孔内，室温避光显色10分钟。
6. 每孔加入终止液50μl混匀终止反应。
7. 酶标仪读值（450nm波长或者450 nm /630 nm波长）。

【结果判断】
用酶标仪单波长450nm或双波长450nm/630nm测定各孔OD值（用单波长测定时需设空白对照一
孔）。
临界值＝阴性（N）OD值×2.1
样本OD值＜临界值为阴性，样本OD值≥临界值为阳性。

【注意事项】
1. 不同批号试剂请勿混用。
2. 反应温度和时间必须严格控制。
3. 严格按说明书操作。

18 DIDRICKSON DRIVE, TORONTO, ONTARIO, CANADA (M2P 1J6)

WEBSITE: HTTP://WWW.UTBIOMED.CA E-MAIL: SUPPORT@UTBIOMED.CA
 UT-BioMED™

Influenza Virus Type B Test (ELISA)

This kit is based on double antibody sandwich assay for the detection of antigen of
influenza Bvirus. Operation process is one step method. The monoclonal antibody against
influenza B virus is coated on the microtiter strips. Add the specimens and enzyme-antibody
conjugate into each well, incubate, wash and add substrate. Read value and evaluate after
stopping.

I. Kit contents: (48 tests)

Microtiter test strips: 12×4 strips
Conjugate: 3.0ml×1 bottle
Positive control: 0.5ml×1tube
Negative control: 0.5ml×1 tube
Washing solution concentrate: 20ml×1 bottle
Substrate A: 3.0ml×1 bottle
Substrate B: 3.0ml×1 bottle
Stopping solution: 3.0ml×1bottle
Sealing film: two Bag sealing: one

II. Storage and expiry:

Store at 2-8℃; Kits are valid for 9 months.

III. Operation protocol:

1. Preparation before test: Take out the kit before test, balance at 18～25; dilute the
washing solution ℃concentrate(1:25) with double distilled water.
2. Turn a cotton swab behind the soft palate upwards to the nasopharynx and take a
smear, dilute the cotton swab in the 0.5ml sterile normal saline in the bottle and mix
thoroughly. Keep close to the bottle and squeeze it. Dispose the swab properly.
3. Take 50μl specimen and negative or positive control into the well respectively. Then
add 50μl conjugate, mix thoroughly. Incubate 30 minutes at 37.℃
4. Wash the strips 5 times and dry by tapping the microtest plate on a paper towel.
5. Add 50μl substrate A and B into reaction wells respectively. Incubate for 10 minutes
at room temperature, avoiding light.
6. Add 50μl stopping solution into each well.
7. Read optical density at 450nm or 450nm/630nm with Microplate reader.
IV. Evaluation:
Measure the absorbance at 450nm or 450nm/630nm with Microplate reader. (One blank
control should be set up at 450nm)
Critical value＝Negative OD value×2.1
Negative: OD Value of specimen＜Critical value;
Positive: OD Value of specimen ≥Critical value.

V. Note:
1. Do not mix reagents from different kit lots.
2. Reaction time and temperature should be strictly controlled.
3. Operate in strict accordance with the protocol.

18 DIDRICKSON DRIVE, TORONTO, ONTARIO, CANADA (M2P 1J6)

WEBSITE: HTTP://WWW.UTBIOMED.CA E-MAIL: SUPPORT@UTBIOMED.CA