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乙脑ELISA检测试剂盒 品牌美国NOVABIOS

乙脑ELISA检测试剂盒 品牌美国NOVABIOS

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乙脑ELISA检测试剂盒 品牌美国NOVABIOS用于暴露于日本脑炎病毒(JEV)是用于检测人血清中的IgE抗体对JEV衍生的重组抗原(JERA)(1-4)的ELISA测定系统。这个测试是为了帮助诊断人类暴露于日本脑炎病毒(JEV)。它不是为了筛选血液或血液成分,仅用于专业的体外诊断用途。该试剂盒尚未针对疫苗诱导的血清学转换研究进行优化。

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  乙脑ELISA检测试剂盒 品牌美国NOVABIOS

 

NOVABIOS JE DetectTM 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.

      乙脑ELISA检测试剂盒 品牌美国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 37oC 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 37oC 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 37oC 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.
  • 更具体的说明请根据以下信息

     

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