Assay human papillomavirus

A CL ISH assay for the detection of human papillomavirus (HPV) DNA was developed, in which the hybridization reaction was performed using either digoxigenin-, biotin-, or fluorescein-labeled probes [64], The hybrids were visualized using AP as the enzyme label and a highly sensitive 1,2-dioxetane phosphate as chemiluminescent substrate. This assay was applied to biopsy specimens from different pathologies associated with HPV, which had previously proved positive for HPV DNA by polymerase chain reaction (PCR). The analytical sensitivity was assessed using samples of HeLa and CaSki cell lines, whose content in HPV DNA is known (10-50 copies of HPV 18 DNA in HeLa cells and 400-600 copies... 

C4. Coutlee, F., Provencher, D., and Voyer, H., Detection of human papillomavirus DNA in cervical lavage specimens by a nonisotopic consensus PCR assay. J. Clin. Microbiol. 33, 1973-1978 (1995). 

To date, the most successful antiviral targets have been directed against viral-specific enzymes. Therefore, the ATPase and helicase activities of the El protein are attractive targets. Papillomavirus DNA replication may also be inhibited by compounds that interfere with the ability of El to bind DNA or to interact with the E2 protein. This chapter will describe a method to assay for specific El DNA binding and cooperative origin binding with the E2 protein and a method to transiently assay papillomavirus DNA replication. The methods described are for bovine papillomavirus type 1 (BPV-1), which has been the molecular prototype of the papillomaviruses. However, the methods can easily be adapted to assay for human papillomavirus replication. 

Quantitatively assessing oncoproteins in HPV-transformed cells is useful for evaluating viral activity and the potential effectiveness of antiviral therapy (4). The following describes immunodetection methods for assaying levels of human papillomavirus E6 and E7 protein in total protein cell lysates. This pro-... 

Kornegay JR, Shepard AP, Hankins C, Franco E, Lapointe N, Richardson H, et al. Nonisotopic detection of human papillomavirus DNA in clinical specimens using a consensus PCR and a generic probe mix in an enzyme-linked immunosorbent assay format. J Clin Microbiol 2001 39 3530-6. 

S., Feindt, FL, Niedbala, R. S., and Tanke, H. (2001) Use of up-converting phosphor reporters in lateral-flow assays to detect specific nucleic acid sequences a rapid, sensitive DNA test to identify human papillomavirus type 16 infection. Clin Chem. 47, 1885-1893... 

Another manifestation of genetic instability is an abnormal number of centro-somes. Centrosomes can be stained and quantified in an assay using anti-centrin antibodies. Both, genetic instability and loss of genomic integrity, are often posttransformation events. However, under certain circumstances they can contribute to cellular transformation directly. An example is the transformation by the human papillomavirus (HPV) E7 protein. The HPV E7 and E6 proteins are the two HPV oncogenes found in 99.3% of all cervical cancers. 

Assay DNA-binding domain of human papillomavirus E2 protein... 

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