Rapid Test for Identification of a Human Papillomavirus 16 E6 L83V Variant

Alemi, Mansour; Andersson, Sonja; Sällström, Johan; Wilander, Erik

doi:10.1097/00019606-199906000-00006

Cited by 13 publications

(12 citation statements)

References 17 publications

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“…To distinguish monoclonal PCR products from polyclonal, single-stranded conformation polymorphism (SSCP), analysis was performed as reported elsewhere (18). In one case for which a clonal PCR product was lacking in the diagnostic tumor (Case 33), further analyses were carried out using Southern blotting and hybridization with a J H probe to determine clonality of the IGH gene as previously detailed (19).…”

Section: Analyses Of Clonal Relationshipmentioning

confidence: 99%

Chromosomal Imbalances in Diffuse Large B-Cell Lymphoma Detected by Comparative Genomic Hybridization

et al. 2002

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Non-Hodgkin lymphomas (NHL) constitute a heterogeneous group of lymphoid neoplasms that display different morphological, immunological, cytogenetic, and molecular genetic features. The revised European-American classification of lymphoma (1) takes all these features into consideration for the distinction of different lymphoma entities. Lymphomas have been associated with a wide range of recurrent chromosomal abnormalities. Some of these can occur as a single cytogenetic alteration and may sometimes be recurrent in a lymphoma entity, such as the t(14;18)(q32;q21), which is found in 85 to 90% of follicular lymphomas. The occurrence of distinct chromosomal changes in specific histopathological subtypes of NHL has improved the understanding of the genetic basis of lymphomagenesis (2). Diffuse large B-cell lymphomas (DLBCL) constitute approximately 40% of all lymphomas and are typically characterized by an aggressive behavior in

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Section: Analyses Of Clonal Relationshipmentioning

confidence: 99%

Chromosomal Imbalances in Diffuse Large B-Cell Lymphoma Detected by Comparative Genomic Hybridization

et al. 2002

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“…The PCR product was run on a SSCP gel and a polymorphism of E6 codon 83 was identified after silver staining by discrepant band pattern of the two sequence variants. The method has been described in detail before (Alemi et al, 1999). The primer pairs used for the HPV analyses are specified in Table 1.…”

Section: Hpv-testingmentioning

confidence: 99%

“…In the present investigation, a collection of invasive cervical adenocarcinomas infected with HPV16 was analysed for the presence of a point mutation in codon 83 of the E6 gene, by a novel, rapid, PCR-based method (Alemi et al, 1999). It was observed that the HPV16 E6 variant was more predominant in the invasive squamous lesions than in the adenocarcinomas.…”

mentioning

confidence: 91%

Uneven distribution of HPV 16 E6 prototype and variant (L83V) oncoprotein in cervical neoplastic lesions

et al. 2000

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Summary A previous Swedish study revealed that both prototype and variant HPV16 E6 oncoprotein, occur in about equal numbers in highgrade cervical intraepithelial neoplasia (HCIN), whereas variant HPV16 predominates in invasive cervical squamous carcinoma. Most of the malignant HPV16 variants contain a common mutation, L83V, in the E6 oncoprotein. In the present investigation, 28 HPV16 positive, invasive cervical adenocarcinomas were collected from a total number of 131 adenocarcinomas. These HPV16-positive cases were evaluated with analysis of the E6 gene, using a recently described PCR-SSCP method for identification of the specific mutation (L83V) in the E6 gene. The results obtained were correlated to findings in 103 preinvasive, HCIN, and 31 invasive cervical squamous carcinomas also infected with HPV16. The HPV16 E6 variant L83V was present in 40% of the HCIN lesions, in 54% of the invasive adenocarcinomas, in comparison to 81% of the invasive squamous carcinomas. The difference between HCIN and squamous carcinomas was statistically significant, P < 0.001, whereas the difference between HCIN and invasive adenocarcinomas was not statistically significant, P = 0.604. Prototype HPV16 and its E6 variant L83V are both prevalent in preinvasive and invasive cervical lesions in Swedish women. However, the obvious predominance of HPV16 variant in squamous carcinomas was not seen in adenocarcinomas. A single amino-acid shift in the HPV16 E6 gene appears to result in a different transforming potential in squamous and glandular cervical lesions.

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“…2 Recently, it has also been used for studies of viral evolution and molecular epidemiology. 6,42 As described for other systems, both types of gels, with and without glycerol, were necessary for detection of all substitutions.…”

Section: Discussionmentioning

confidence: 99%

Application of Single-Strand Conformation Polymorphism to the Study of Bovine Viral Diarrhea Virus Isolates

Jones¹,

Weber²

2001

J VET Diagn Invest

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Abstract. Single-strand conformation polymorphism (SSCP) analysis of polymerase chain reaction (PCR) products is a genetic screening technique for rapid detection of nucleotide substitutions in PCR-amplified genomic DNA or cDNA. It is based on the observation that partially formamide-denatured double-stranded DNA migrates as 2 single-stranded DNA molecules when electrophoresed in nondenaturing polyacrylamide gels. The mobility depends on the 3-dimensional conformation of the strand under the conditions used. It is possible to discriminate between DNA strands differing in only 1 nucleotide. The method was applied to the analysis of Bovine Viral Diarrhea Virus (BVDV) isolates. Reference and Argentinian strains were assessed for variations in their 5Ј untranslated region (5Ј-UTR). The PCR products of the 5Ј-UTR ends were formamide denatured and compared by SSCP analysis in nondenaturing 15% polyacrylamide and 15% polyacrilamide-5% glycerol gels. The reference strains SD-1, Singer, and Oregon C24V had differences in electrophoretic patterns. Despite the high conservation among the 5Ј-UTR of pestiviruses, the method allowed discrimination among all 9 Argentinian isolates. The 5Ј-UTR of a fetal kidney-derived isolate (1R93) was PCR amplified and cloned in a plasmid vector; the SSCP analysis of 30 PCR products obtained by direct amplification over randomly selected clones produced 5 different banding patterns, indicating the existence of viral quasispecies. The results show that SSCP may be used to identify and differentiate among BVDV isolates.

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Rapid Test for Identification of a Human Papillomavirus 16 E6 L83V Variant

Cited by 13 publications

References 17 publications

Chromosomal Imbalances in Diffuse Large B-Cell Lymphoma Detected by Comparative Genomic Hybridization

Chromosomal Imbalances in Diffuse Large B-Cell Lymphoma Detected by Comparative Genomic Hybridization

Uneven distribution of HPV 16 E6 prototype and variant (L83V) oncoprotein in cervical neoplastic lesions

Application of Single-Strand Conformation Polymorphism to the Study of Bovine Viral Diarrhea Virus Isolates

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