How to deal with errors in the results of ELISA kit experiments - Database & Sql Blog Articles

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The problem of the ELISA kit results in the "flower plate" has a great impact on the test data, and often causes the tester to make a false judgment, which must be paid attention to and avoided in the experiment.

In response to this "flower board" phenomenon, we have summarized some experiences through analysis and judgment of many experiments, and analyzed as follows:

1 specimens, reagents, instruments

1.1 Blood specimens of specimen entry and exit personnel.

1.2 reagents HBsAg, HCV, HIV, syphilis reagents are ELISA kits from different manufacturers.

1.3 instrument LD4-2 brain heart machine BIDRAD1575 type washing machine BIDRAD550 microplate reader.

2 methods

Blood samples of all inspected and entry personnel shall be tested in accordance with the instructions of the ELISA kit provided by each manufacturer for HBsAg-HCV-HIV syphilis. All reagents are used within the validity period.

3 results

The "flower board" problem sometimes occurs with reagents from different manufacturers.

4 analysis

In the process of testing all specimens, the instruments used and the loading process are in a normal state, and the steps are performed according to the requirements of various reagent instructions, excluding the influence of the instruments and operating procedures used in the experiment on the experimental results. Blank, negative and positive controls and indoor quality control values ​​in the ELISA kit were normal, and only the blood samples tested were abnormal. According to the fact that the ratio of positive rate reported by the State and the Center for Disease Control and Prevention should not be very high, HBsAg has been tested by rapid gold standard reagent before ELISA, and the positive specimen has been controlled. Therefore, the cause may be problems in the processing of specimens, particularly placement time, centrifugation speed and centrifugation time. Therefore, the processing of specimens becomes the main suspicious factor of the experimental results.

In order to avoid the occurrence of "flower board" phenomenon, reduce the false positive rate of detection, and obtain accurate experimental results, ELISA reagents with high sensitivity, good specificity, good stability, small batch difference, convenient operation and time saving should be selected.

Using the ELISA kit for blood test results of HBsAg-HCV-HIV and syphilis, blank and negative, positive control and indoor quality control hole results were normal, according to the data provided by the Ministry of Health Clinical Inspection Center, HBsAg0.5ng1ng-HCV- The normal range of HIV syphilis is 2NCU, but the OD value of blood sample wells is relatively high (most OD values ​​≥0.070). The positive rate of the specimens is very high. In addition to the control wells, each plate is sometimes as high as 10 wells, and most of the true positive specimens have weak positive results.

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