CRISPR/LAMP Microfluidics for Human Monkeypox Diagnosis

Human monkeypox is an infectious disease that has recently become a global public health threat. Monkeypox (MPX) virus is the causative agent of the disease. It is a zoonotic disease endemic to the African continent, which has existed for many years. However, recently, cases have been encountered in many non-endemic countries at the global level. It has been determined as a moderate epidemic risk by the World Health Organization (WHO). Although the number of cases in our country is low, the high number of cases in countries with social and commercial networks pose a threat to our country. It is essential to take precautions against the possibility of a possible epidemic. Since the epidemic affects large masses, strategies to prevent, keep under control, and improve the epidemic must be systematically designed. For the successful management of health strategies, the first step is to make an accurate and reliable diagnosis. With this project, we aim to develop a new point-of-care (PoC) diagnostic test with high specificity and accuracy that can be used for the diagnosis of human monkeypox. We aimed to develop a disposable microfluidic chip that can be used for the detection of monkeypox virus by Clustered regularly spaced short palindromic repeats (CRISPR) associated (CRISPR-Cas) combined loop-mediated isothermal amplification (LAMP) technique (CRISPR/LAMP). CRISPR/LAMP technology is a new generation combined molecular method with high accuracy and specificity that can be designed pathogen-specifically. The fact that the experimental process includes multiple stages brings with it difficulties such as the risk of contamination, the need for specialist personnel and the need for a laboratory environment. We aim to develop a new diagnostic test at a full PoC level that takes advantage of the microfluidic chip technology and the CRISPR/LAMP method and eliminates its difficulties. In project scope; I) design-production of the microfluidic chip, II) lyophilization of the reaction components on the microfluidic chip, III) development of the optical reading system, IV) performing the CRISPR/LAMP reactions on the chip and monitoring the results. For the detection of human monkeypox, we envisage that we will be able to develop a disposable, fast, practical and highly accurate new generation PoC molecular diagnostic test, where the results can be monitored with an application on a smartphone. The techniques included in the PoC diagnostic test that we plan to develop are unique and this type of diagnostic test will be created for the first time with this study.