| Background and Objection: Nowadays the close relationship between human papillomavirus and cervical cancer was incontestable fact. Human papillomavirus infection, especially high-risk human papillomavirus infection was the major cause of cervical cancer and its precancerous diseases. HPV infection strongly indicated the possibility of squamous intraepithelial lesions (SIL). Recent studies had shown that high-risk HPV was present in more than 99.7% of cervical carcinomas worldwide. The detection and genotyping of HPV was an important evidence for the dealing with the cervical diseases, and also an indispensable context of the screening of cervical cancer. However, conventional HPV detecting methods (such as cytology, PCR, Southern blot) had many disadvantages such as high pseudopositive rate, high-cost, inefficenct, complicated protocols and so on, all these disadvantages limited their use in clinic. So more sensitive, reliable, efficient and low-cost methods than conventional testings should be developed as screening tools for the detection of HPV. DNA microarray was a jumped-up technique in 90's 20th century that contains molecular biology, material science, microelectronics, physics, chemistry, computer and information science and so on. DNA chip had became the study focus now. Geneanalysis can be realized high-through, rapidly, sensitively and efficient in many samples by DNA microarray. So DNA microarray was widely used in DNA sequencing, the analysis of gene expression, genome, gene diagnosis, the study and explore of new medicines. Recently, DNA chip was widely used in the gene diagnosis of microorganism following the rapid development of the technique and it brought technical revolution in the gene diagnosis of microorganism. We performed HPV detection and genotyping with the newly designed DNA chip. In this study, we put DNA microarray into the detection and genotyping of HPV and constructed the DNA chip and gave a primary study on the protocols of HPV DNA microarray that contained the most common HPV types(HPV6,11,16,18) and ascertained its best using conditions.Materials and methods: Collected paraffin embeded tissue specimens of condyloma acuminata (50 samples) and cervical cancer ( 22 samples). Isolated DNA by the method of hydroxybenzene-chloroform extraction. Tested HPV by HPV-PCR detection kit and purified the positive samples. Made plasmids and detected it by DNA sequencing, blasted in GenBank and saved the plasmids that contained HPV L1 region genome of HPV6, HPV11, HPV16, HPV18. One common oligonucleotide probe to HPV and four specific oligonucleotide probes to their genotypes (HPV6, 11, 16, 18) were designed by the characters of the genome of HPV L1 region. The non-homology sequences of vegetative genes were used as negative control, one HBV gene fragment as positive control, pointing solution that contained no gene fragment as blank control. All the probes were pointed on glass slides by GMS 417 Arrayer. Amplified plasmids wer.e labeled by PCR fluorescence(CY5)-primer and CY5-dUTP. The labeled plasmids were applied to the DNA chip. The hybridization was performed at scalar temperatures (43 C, 45 C, 47 C, 49 C, 51C) for one hour. After hybridization the chips were scanned by GMS418 Array Scanner. The signals were collected by ScanArray. The difference of the signals under different labeling methods and hybridization temperatures was analysed by SPSS 10.0 for Windows. Comparison of proportions of the tow groups was performed by t-test (a=0.05).Results:1. The plasmids constructed from the positive HPV PCR products were confirmed as HPV6, HPV11, HPV16 and HPV18 by DNA sequencing.2. The efficiency of labeling by PCR fluorescence(CY5)-primer was higher than that of labeling by CY5-dUTP. The difference between the tow groups was significant (P<0.01) in this DNA chip.3. When hybridization temperature was 43 C or 45 C, there was mis-matched signals. When hybridization temperature was 51C, there was no hybridization signal. When hybridization temperature was 47 C or 49 C, there w...
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