Diagnosis of Human Papillomavirus Using In Situ Hybridization and In Situ Polymerase Chain Reaction View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2001-09-17

AUTHORS

Marilena Aquino de Muro , Ralph Rapley , Gerard J. Nuovo

ABSTRACT

In situ hybridization is the only DNA—or RNA-based molecular biology based test that allows for the direct correlation of the results with the histologic and cytologic features of the sample. The DNA/RNA extraction that precedes filter hybridization (hybrid capture or Southern blot) hybridization and the polymerase chain reaction (PCR) precludes this type of analysis. In order for the in situ hybridization to detect a given DNA or RNA target, there must be at least 10–20 copies of the target per cell. In comparison, Southern blot hybridization and the hybrid capture assay can detect one DNA or RNA target per 100 cells. PCR is even more sensitive if the hot start maneuver is employed; under these conditions, it can detect one DNA or RNA target per 100,000 cells. It is evident that in situ hybridization is a relatively insensitive test. A reflection of this relative insensitivity is seen in occult or latent infection by a virus where the copy number is low. In such situations, the virus is rarely detected by in situ hybridization even though it was detected by either PCR or filter hybridization (1–6). As stressed in this chapter, this is actually an advantage of in situ hybridization over the more sensitive assays. In situ hybridization detects only productive infection by human papillomavirus (HPV). It will not detect the low copy subclinical or “incidental” infection of HPV in the setting of a normal Pap smear (Fig. 1). When one realizes that incidental HPV infection can occur in 15–20% of normal Papanicolaou (Pap) smears, as determined by the hybrid capture or PCR assays, it is evident that it is advantageous for in situ hybridization NOT to detect the virus, given the realization that most concur there is no reason to clinically treat HPV in the absence of any pathology. Fig. 1. Detection of HPV DNA in Pap smears. (A) A Pap smear that was within normal limits; HPV DNA was detected using the hybrid capture system (not shown). However, HPV DNA was not detected by in situ hybridization (A, pink is negative and blue is positive). (B) Specimen is from a woman with a Pap smear diagnosis of ASCUS. HPV DNA was detected by the hybrid capture system and in situ hybridization (B). The in situ test detects only HPV associated with productive infection and, thus, clinical disease. More... »

PAGES

113-136

References to SciGraph publications

Book

TITLE

Gene Probes

ISBN

1-59259-238-4

Identifiers

URI

http://scigraph.springernature.com/pub.10.1385/1-59259-238-4:113

DOI

http://dx.doi.org/10.1385/1-59259-238-4:113

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1007669136


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