Cihan University-Erbil Conferences, 4th International Conference on Biological & Health Sciences (CIC-BIOHS’2022)

Font Size: 
Development and validation of NLRP3 immunochromatography assay as an indicator for Inflammation and Bad Seminal Quality
Nuha Abdulsalam Younis, Haider Faisal Ghazi, Ula Mohammed Alkawaz

Last modified: 2022-08-30

Abstract


Estimation of seminal inflammation is highly necessary for management of Infertile patients, unfortunately peroxidase staining and leukocyte count is not indicative for true inflammation. This study aimed to develop a rapid NLRP3 immunochromotography (ICT) strip as an alternative tool for indication of inflammation rather than confusing Endtz test. Methodology: Immunochromotography cassette developed in the Department of Microbiology/ College of Medicine/ AL-Nahrain University. Polyclonal capture rabbit anti-NLRP3 (test line) and goat anti-rabbit IgG antibodies (control line) were applied on each nitrocellulose membrane. Gold-in-a-BoxTM conjugation kit (40nm. 15OD) conjugated anti-NLRP3 were adsorbed in conjugate pad, then assembled in the test cassette. The developed kit was tested in vitro using ELISA kit depending on serial concentration of standard NLRP3 protein. Fifty infertile patients included in this study, seminal fluid analysis and Endtz test were done. Results: the developed NLRP3 ICT was sensitive at the level of 5 ng/ml. NLRP3 was detected in seminal fluid of 30 (60%) of infertile patients. ICT assay showed 92.86 sensitivity and 81.82 specificity in comparison with ELISA test. Conclusion: NLRP3 protein in seminal plasma may be associated with the increased leukocytospermia, bad sperm quality among infertile male with higher sensitivity than Endtz test. NLRP ICT kit could be used for diagnosis of inflammation in infertile men.

Keywords—Leukocytospermia, Immunochromatography, Infertility, Inflammasome.

 

DOI: http://doi.org/10.24086/biohs2022/paper.757


Full Text: XML