乙腦ELISA檢測試劑盒 品牌美國NOVABIOS用于暴露于日本腦炎病毒（JEV）是用于檢測人血清中的IgE抗體對JEV衍生的重組抗原（JERA）（1-4）的ELISA測定系統(tǒng)。這個測試是為了幫助診斷人類暴露于日本腦炎病毒（JEV）。它不是為了篩選血液或血液成分，僅用于專業(yè)的體外診斷用途。該試劑盒尚未針對疫苗誘導(dǎo)的血清學(xué)轉(zhuǎn)換研究進(jìn)行優(yōu)化。

  乙腦ELISA檢測試劑盒 品牌美國NOVABIOS

 

NOVABIOS JE Detect^TM IgM ANTIBODY

CAPTURE ELISA (MAC-ELISA)

• Positive, negative and unknown serum to be tested should be assayed in duplicate. Refer to flow chart at the end of this section for illustration of this procedure. Twenty-two test specimens can be tested in duplicate on one 96 well plate.

      乙腦ELISA檢測試劑盒 品牌美國NOVABIOS

• Mark the microtitration strips to be used.
• Dilute test sera, and controls to 1/100 using the provided Sample diluent. Use small polypropylene tubes for these dilutions and at least 4 µL of sera and positive and negative controls. For example: 4 µL serum plus 396 µL of Sample Dilution Buffer for JE Detect IgM to make 1/100 dilution.2

       

• Apply the 0 µL/well of 1/100 diluted test sera, JE Detect Negative Control, and JE Detect IgM Positive Control to the plate by single or multi-pipetter as appropriate. An exemplary arrangement for twenty-two test serum samples in duplicate is shown below.

Example for Serum Sample Application

 

1

2

3

4

5

6

7

8

9

10

11

12

A

JE Negative

S#1

S#3

S#5

S#7

S#9

S#11

S#13

S#15

S#17

S#19

S#21

Control.

B

JE Negative

S#1

S#3

S#5

S#7

S#9

S#11

S#13

S#15

S#17

S#19

S#21

Control.

C

JE IgM

S#2

S#4

S#6

S#8

S#10

S#12

S#14

S#16

S#18

S#20

S#22

Positive

Control.

D

JE IgM

S#2

S#4

S#6

S#8

S#10

S#12

S#14

S#16

S#18

S#20

S#22

Positive

Control.

E

JE IgM

S#2

S#4

S#6

S#8

S#10

S#12

S#14

S#16

S#18

S#20

S#22

Positive

Control.

F

JE IgM

S#2

S#4

S#6

S#8

S#10

S#12

S#14

S#16

S#18

S#20

S#22

Positive

Control.

G

JE Negative

S#1

S#3

S#5

S#7

S#9

S#11

S#13

S#15

S#17

S#19

S#21

Control.

H

JE Negative

S#1

S#3

S#5

S#7

S#9

S#11

S#13

S#15

S#17

S#19

S#21

Control.

 

• Cover the plate with parafilm just on the well opening surface, so the bottom of the plates is not covered.

 

Note: This is to make sure the temperature distribution is evenly spread out in all wells from bottom and sides; any extra parafilm can be cut-out once the top is sealed to block evaporation.

 

 

• Incubate the plate at 37^oC for 1hour in a humidified incubator with water container. Humidification can be achieved using a water tray at the bottom of incubator.

 

Note: Do not stack plates on top of each other. They should be spread out as a single layer. This is very important for even temperature distribution. Do not use CO₂ or other gases. Do not place plates in contact with any wet substances such as wet paper towels etc.

 

CORRECT METHOD

 

 

• After the incubation, wash the plate 6 times with automatic plate washer using 1x Wash buffer. Use 300 µl per well in each wash cycle.

 

• Add 50µl /well of JERA into row A-D and 50µl /well of

NCA into row E-H by multi-pipetter.

An exemplary application for JERA and NCA is shown below.Example for JE Antigens Application

1

2

3

4

5

6

7

8

9

10

11

12

A

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

B

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

C

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

D

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

JERA

E

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

F

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

G

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

H

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

NCA

 

• Cover the plate with parafilm just on the well opening surface, so the bottom of the plate should not be covered (see step 5).

 

• Incubate the plate at 37^oC for 1 hour in a humidified incubator with a water container for humidification (see step 6).
• After the incubation, wash the plate 6 times with automatic plate washer using 1x Wash buffer. Use 300 µl per well in each wash cycle.
• Add 50µl /well of ready to use Enzyme-HRP conjugate into all wells by multi-pipetter.
• Cover the plate with parafilm just on the well opening surface, so the bottom of the plate should not be covered (see step 5).

 

• Incubate the plate at 37^oC for 1hour in a humidified incubator (see step 6).

 

• After the incubation, wash the plate 6 times with automatic plate washer using 1x Wash buffer.
• Add 150µl /well of EnWash into all wells by multi-pipetter.
• Incubate the plate at room temperature for 5 minutes without any cover on the plate.
• After the incubation, wash the plate 6 times with automatic plate washer using 1x Wash buffer.
• Add 75µl /well of Liquid TMB substrate into all wells 3

multi-pipetter.

• Place and incubate the plate at room temperature in a dark place (or container) for 10 minutes without any cover on the plate.
•  
• After the incubation, add 50µl /well of Stop solution into all wells by multi-pipetter and incubate at room temperature for 1 minute without any cover on the plate.
• After the incubation, read the OD 450 value with a Microplate reader.

• 更具體的說明請根據(jù)以下信息

   

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