Our team proposes a kit that can be used for mass screening of high risk Human Papillomavirus. This can prevent many cases of HPV infection from progressing to cervical cancer by early detection and preventative measures.We propose a kit that can work in a low-resource setting, at room temperature and without the use of sophisticated laboratory equipment or highly trained personnel. Our kit comprises of three major steps. (For more details regarding the kit workflow, please see the Solution page under the Project tab)We could only perform the wetlab experiments for the last two steps of our kit workflow due to biosafety restrictions. However, the first two steps of the workflow are routinely done for similar systems. Especially the DNA extraction step is commonly and commercially available [1].As proof of concept, we were able to successfully demonstrate a positive HPV on a lateral flow assay based dipstick in the form of two solid lines. This result was significantly distinguishable from a negative HPV wherein only the test line (Line closer to the sample pad) could be observed.

These results could also be successfully replicated multiple times strengthening our hypothesis. The same results could also be obtained at room temperature by simply increasing the incubation time to 4 hours.
Our system can detect a very low viral DNA load of about 0.15 picomoles. This can be said on the basis of the fact that all of our successful experiment replicates have been carried out using only 3uL of 50nM E7.

 Three Guide RNAs with E7 present

For more details of the same, please check out our Results page under the Wetlab Tab.Thus, we can successfully conclude that it is possible to detect the E7 gene of HPV16 at room temperature only utilizing basic laboratory techniques and equipment. This nucleic acid detection method would also lead to increased sensitivity and accuracy when compared with other methods of detection like antigen testing.

All three guide RNAs incubated at Room Temperature with the negative control