Generally this is a multi-step process. You do your first tests with leaves inoculated and collected at optimum disease conditions along with control uninoculated leaves. If results are promising, you include leaves inoculated with pathogens that might be confused with your target pathogen. If results are still promising, you ask collaborators to send you naturally infected leaves you can test in conjunction with your positive and negative controls. If you have some sort of extension service, an extension agent could help you with the last step. If not, you can contact grower groups and they are generally delighted to help develop good diagnostic tests.
It depend on the type of disease you are studying. Nevertheless, it is possible to collect a group of leaves from each plant. Let us assume twenty-five sheets and then make a disease scale consisting of four or five degrees depending on the number of spots on each leaf and distribute the leaves on the degrees of the scale and then use the formula of McKinney to calculate disease severity.