Point-of-care testing (POCT) technology, using lateral flow assays and microfluidic systems, facilitates cost-effective diagnosis, timely treatment, ongoing monitoring, and prevention of life-threatening outcomes. Aside from significant advancements demonstrated in academic research, implementation in real-world applications remains frustratingly limited. The divergence between academic developments and practical utility is often due to factors such as operational complexity, low sensitivity and the need for trained personnel. Taking this into consideration, our objective is to present a critical and objective overview of the latest advancements in fully integrated one-step POCT assays for home-testing which would be commercially viable. In particular, aspects of signal amplification, assay design modification, and sample preparation are critically evaluated and their features and medical applications along with future perspective and challenges with respect to minimal user intervention are summarized. Associated with and very important for the one-step POCT realization are also readout devices and fabrication processes. Critical analysis of available and useful technologies are presented in the SI section.