The sensitivity of in situ hybridization for detection of human papillomavirus

We studied a sensitivity of HPV DNA detection by in situ hybridization method using 3H labeled HPV DNA. The materials were CaSki cells and SiHa cells which were derived from as a negative control. The total cellular DNAs extracted from these cell lines were estimated copy numbers of HPV 16 DNA using Southern blot hybridization. In our result, CaSki cell has 400 copies/cell, SiHa cell were appeared to have 1-5 copies/cell. Simultaneously these cells were fixed by periodate-buffered lysine-paraformaldehyde-glutaraldehyde (PLPG) and were detected HPV 16 DNA using in situ hybridization. We detected HPV 16 DNA in CaSki cells and SiHa cells by in situ hybridization also. We concluded that the sensitivity of our in situ hybridization technique is 1-5 copies/cell.