Researcher ORCID Identifier
Date Degree Awarded
Open Access Dissertation
PHD in Applied Life Sciences
First Thesis/Dissertation Advisor
Sexually transmitted infections caused by Chlamydia trachomatous (CT) and Neisseria gonorrhea (NG) are often under-diagnosed, miss-diagnosed, and not properly treated, leading to long term complications such as ectopic pregnancy and infertility, and to emergence of drug resistance. Dengue virus (DENV) infections can cause dengue fever (DF), which if not properly managed can lead to Dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS), causing significant morbidity and mortality. In both cases, early and accurate detection at the point of care is critical to facilitate proper patient care and disease management. Nucleic acid amplification tests (NAATs) provide suitable sensitivity and specificity, but are often complex, expensive, and difficult to implement at the point of care, especially in low resource settings. We have developed a prototype system consisting of a cartridge and compact instrument that can execute sample preparation, isothermal Loop Mediated amplification (LAMP) and lateral flow detection of these pathogens. For CT and NG, we have established singleplex and duplex LAMP assays, in liquid and paper form, with lateral flow detection. We have performed inhibition testing, demonstrated full process execution outside the cartridge, then implemented the 4 process in the cartridge and device, demonstrating detection down to 100 Elementary Bodies (EB)/mL for CT and 100 colony forming unit (CFU)/mL NG in urine. For DENV, we established a pan-serotype singleplex RT-LAMP assay and a duplex assay with MS2 as an internal amplification control, in liquid and paper form. We demonstrated that the master-mix in paper form is stable upon storage at RT and 40oC over 8 weeks. For sample preparation starting from whole blood, we performed plasma separation coupled to amplification, with inhibitor testing. The next step is a front to back experiment and implementation in a modified cartridge for the detection of all DENV serotypes. The herein described assay processes and prototype systems can be further developed to enable point of care diagnosis for these and other pathogens in low resource settings.
©2021 Abrar N Al Maghribi
Al Maghribi, Abrar. (2021). POINT OF CARE INFECTIOUS DISEASE DIAGNOSIS VIA ISOTHERMAL NUCLEIC ACID AMPLIFICATION INTEGRATED INTO A SAMPLE TO ANSWER DEVICE. KGI Theses and Dissertations, 17. https://scholarship.claremont.edu/kgi__theses/17.