CA199  Tumor Markers

INTENDED USE

Detection Kit for Carbohydrate antigen 19-9(CA199)(ELISA). It is used in quantitative tests for CA19-9 in human serum.

SUMMARYANDCLINICALSIGNIFICANCE

Carbohydrate antigen 19-9 (CA19-9) is a tumor-associated antigen oligosaccharide and also called pancreatic cancer antigen or sialylated Lewis antigen. CA19-9 is a molecular weight over 5000, 000 and its epitopes is formed of glycoproteins with mucin in serum. CA19-9 present in colorectal cancer cells, and also present in the cancer tissue of pancreas, lung, thyroid, ovary, prostate.

This reagent mainly is used for dynamic monitoring of cancer patients to determine disease progression or auxiliary therapeutic effect, but its concentration is not correlated with tumor size, growth, and the degree of malignancy grading/staging. In some benign diseases (Such as acute pancreatitis, cholecystitis, cholangitis cholestasis, hepatitis, etc.) can also be seen that the index increased in varying degrees. Therefore, the CA19-9 should not be considered an effective general screening test for the general population of cancer screening, early diagnosis and other purposes.

PRINCIPLE

CA-19-9 uses a “sandwich principle”, Enzyme-linked immunological sorbent assay. To measure CA-19-9 levels in serum, plastic wells coated with a monoclonal antibodies of CA-19-9 are supplied in the kit. After the patient’s specimen and another mono-antibody labelled with HRP are added, CA-19-9, if present, is fixed to the solid phase antibody and creating a HRP-antibody—CA-19-9—antibody “sandwich”. After TMB substrate added, the result is obtained by EIAplate reader

PREPARATIONS

1.  Allow all specimens and reagents to reach room temperature and mix thoroughly by gentle inversion before use.

2. Prepare Wash Buffer by diluting Wash Concentrate 20-fold with deionized

ASSAYPROCEDURE:

1.Mark the microtitration strips to be used. All the calibrators and controls should set duplicate.

2.Dispense 50 ul of calibrators/controls/samples into wells.

3.Dispense 50 ul of HRPConjugateto each well. Covered the strips with a plate sealer. Mix it gently by swirling the microtiter plate on flat bench. Incubate the plate at 37°C for 1 hour.

4.Wash each well for 3 times, 10 seconds each time. (See wash procedure).

5.Dispense 50 ul of chromogen Ato each well.

6.Dispense 50 ul of chromogen B to each well.

7.Covered the strips with a fresh plate sealer.

8.Mix it gently by swirling the microtiter plate on flat bench. Incubate the plate at 37°C for 15 minutes.

9.Dispense 50 ul ofstoppingsolutionto each well and mix completely.

10. Readtheabsorbanceof theplate within 10 minutes. (See read procedure)

PRECAUTION FOR USERS

1. Handling should preclude any pipetting by mouth. Use only pipettes with disposable tips for each specimen.

2.Do not mix materials from different master lots. Do not use kit components beyond the expiration date.

3.All materials should be brought to room temperature before use.