Use of Citrus limon Juice as an Alternative Antigen Retrieval Solution for Formalin Fixed Breast Cancer Tissue in Immunohistochemistry

Abstract

Background: The potential of Citrus limon fruit as a source of antigen retrieval agent is a significant step in the locally made brand of antigen retrieval solution with its availability and potential use in research and diagnosis. The cost of Immunohistochemistry tests is on the high side in Nigeria as a developing country, and many cancer patients have lost their lives in the cause of seeking alternative remedies to their prevailing health challenges due to financial constrain encountered in the cost of Immunohistochemistry (IHC) test. Its relevance makes the diagnosis of cancer effective for proper treatment and prognosis. Methods: Fifty (50) positive formalin-fixed breast tissues previously diagnosed using commercially prepared antigen retrieval solution (Thermoscientific – ephredia) were randomly selected for this study. Heat-induced epitope retrieval method (HIER) was used along with breast markers, namely Estrogen receptor (ER), Progesterone receptor (PR), and Human Epidermal Growth Factor Receptor 2 (HER-2) antibodies. Haematoxylin and eosin Techniave were used to demonstrate the general tissue structures to show the pleomorphic appearance of the cells invading the stroma, revealing an irregular shape with hyperchromatic nuclei. Results: Nuclear and cytoplasmic meshwork exposed without any background stain was observed. Estrogen receptor marker results in C. limon juice antigen retrieval was positive with a brown intensity score of 3 and proportion of stain score 5, having Thirty five (35) sections positive (70%). Progesterone receptor results in C. limon juice antigen retrieval revealed positive with a brown intensity score of 3 and a proportion of stain 5, with Forty-one (41) sections positive (82%). HER-2 results showed seventeen (17) sections (34%) positive with complete and intense brown circumferential membrane staining within >10% of tumor cells in C. limon juice antigen retrieval. Conclusion: The use of citrus juice is another source of antigen retrieval solution that can be explored for the immunohistochemistry test.