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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
For more details click here

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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2019.8(6): 2638-2647
DOI: https://doi.org/10.20546/ijcmas.2019.806.317


Development of Latex Enhanced Qualitative Immuno Assay for High Sensitivity CRP Detection
Praveena1*, C.V. Raghuveer2 and D.M. Vasudevan3
1Yenepoya university, Deralakatte, Mangalore, Karnataka, India- 575018
2Sri Devaraj Urs Academy of Higher Education & Research, Kolar, Karnataka - 563 101, India
3Agape, Cochin, Kerala - 683562, India
*Corresponding author
Abstract:

High-sensitivity C-reactive protein (hsCRP) is an important marker of inflammation to predict the risk of CVDs. Existing reagent system for the detection of hs CRP has its limitation such as higher cost and requirement of sophisticated instruments, which minimize the utility of this effective marker utility in Indian population. The study was aimed to formulate and validate an improved reagent system for the detection of hs CRP. The study was targeted to formulate a latex-enhanced reagent format by selecting inert microscopic latex particles. Antibody titration for optimal coating was confirmed in different standard conditions. Reaction buffer characterization for colloidal stability was confirmed and sensitivity setting of the reagent was carried out. Performance validation of the assay was carried by confirming sensitivity, specificity, and precision. Studies of Interference substance testing was carried out for hemoglobin and Bilirubin. The prepared reagent system were tested and compared with existing standard hs CRP reagent systems. An effective reagent system was prepared by selecting Thermo fisher latex of size 0.46 μm with functional modification as carboxyl modified latex. Protein optimization trial yield as 500μg- 750μg of Anti CRP required for 1mg weight latex. BSA 1 % w/v showed good colloidal stability at room temperature and at 37ËšC. Performance validation of the assay was carried out and confirmed the sensitivity specificity, precision and interference testing on hemoglobin and Bilirubin. Comparison of the sample with existing reagent systems shows the effectiveness of the reagent system the prepared reagent formulation was rapid, cost effective, and reproducible and was comparable with the reference method used in clinical practice. The advantage of this proposed methodology is that it does not require any high end automated systems for the estimation.


Keywords: Hs CRP, agglutination, Stabilising agent, Latex beads, Colloidal stability
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How to cite this article:

Praveena, C.V. Raghuveer and Vasudevan, D.M. 2019. Development of Latex Enhanced Qualitative Immuno Assay for High Sensitivity CRP Detection.Int.J.Curr.Microbiol.App.Sci. 8(6): 2638-2647. doi: https://doi.org/10.20546/ijcmas.2019.806.317