Making SARS-CoV-2 IgM/IgG controls with patient samples

For quality control, use controls prepared as follows:

1. Negative serum collection: collect serum from those who are not infected with COVID-

19. Each sample should be clear, transparent, and free of hemolysis, lipolysis, or

jaundice. It is recommended to collect a sample volume of not less than 15mL for a

1-month quality control length. Mix the collected sera evenly. If you can not collect

totally 15mL in one day. the serum can be put into 2-8° C for max. 3 days to allow to

get more sample volume.

▪ The IgM negative control has a target value range of COI<0.8

▪ The IgG negative control has a target value range of COI<8

2. Positive serum collection: collect serum from those who are infected with COVID-19

(confirmation). Choose samples that are about 2 to 8 times of the cutoff value (for IgM

with the value of 2-8, for IgG with the value of 20-80). Each sample should be clear,

transparent, and free of hemolysis, lipolysis, or jaundice. It is recommended to collect

a sample volume of not less than 15mL for a 1-month quality control length. Mix the

collected positive sera evenly. It is recommended to inactivate positive sera at 56 ° C

for 30 min. If there are too few positive samples in the required concentration range,

you can use the negative samples to dilute the inactivated positive samples to the

required concentration. (Use negative serum to run 1:100, 1:50, 1:25, 1:10, 1:5, and 1:2

dilution so that you can determine the dilution factor.) It is recommend to confirm

calculated reactivity after dilution.

3. Preparation of quality control: observe the state of pooled negative serum and positive

serum. If it is not clear, it is recommended to centrifuge it at 2500g for 10min. Collect

the supernatant.

4. Aliquot and storage: after the preparation is completed, you can confirm on the

Mindray system whether the prepared quality control meets the requirements. After

confirming that it is correct, aliquot the negative control and the positive quality

control as 200uL / tube, respectively, and place it under -20 ℃ for long-term storage

(28 days).

5. Test: take a tube of quality control out from the fridge. Leave it at room temperature

for about 30 minutes to thaw thoroughly. Mix it and test it on the analyzer. Discard it

according to local regulation after finishing the test.

6. Quality control target value and range establishment: each day before the start of the

patient sample test and after the completion of the patient sample test, test one

negative control and one positive control in duplicate respectively. Run such tests for

5 days and you can obtain 20 results for each level of control. Calculate the mean and

it can be used as the target value.

The negative control has a target value range as the table below.

Negative Control

IgG 0~8 U/mL

IgM 0~0.8

(When the negative control result is within the above range, it is considered acceptable.
 If you would love to use the Levey-Jennings chart (mean±3SD), you can set the mean

as 4 U/mL for IgG and 0.4 for IgM and SD as 1.33 U/mL for IgG and 0.133 for IgM.

You can also set the mean and SD as per your needs and make sure that the range is

within the ones mentioned in the table above.)

Set the SD of the positive control as the table below.

Positive control

IgG Target value (mean) x 15%

IgM Target value (mean) x 15%

7. The target values of the quality control materials are lot specific and target value

assessment as described above has to be performed for every assay lot.

8. In case quality control fails for any reason, thaw a new aliquot and re-assess the

performance of the assay.

9. Controls should be run individually at least once every 24 hours when the test is in

use, once per reagent kit, and following each calibration.

10. Please follow the applicable government regulations and local guidelines for quality

control.