Molecular detection methods of human papillomavirus (HPV)

2009 ◽  
Vol 24 (4) ◽  
pp. 215-222 ◽  
Author(s):  
Apostolos Zaravinos ◽  
Ioannis N. Mammas ◽  
George Sourvinos ◽  
Demetrios A. Spandidos
Keyword(s):  
Human Papillomavirus ◽  
Molecular Detection ◽  
Blot Hybridization ◽  
High Sensitivity ◽  
Detection Methods ◽  
Dot Blot ◽  
Hpv Dna ◽  
Moderate Sensitivity ◽  
Hybridization Assays

Human papillomavirus (HPV) testing can identify women at risk of cervical cancer. Currently, molecular detection methods are the gold standard for identification of HPV. The three categories of molecular assays that are available are based on the detection of HPV DNA and include (1) non-amplified hybridization assays, such as Southern transfer hybridization (STH), dot blot hybridization (DB) and in situ hybridization (ISH); (2) signal amplified hybridization assays, such as hybrid capture assays (HC2); (3) target amplification assays, such as polymerase chain reaction (PCR) and in situ PCR. STH requires large amounts of DNA, is laborious and not reproducible, while ISH has only moderate sensitivity for HPV. The sensitivity of the HC2 assay is similar to that of PCR-based assays, with high sensitivity being achieved by signal rather than target amplification. PCR-based detection is both highly sensitive and specific. Since PCR can be performed on very small amounts of DNA, it is ideal for use on specimens with low DNA content. In the future, with the advance of technology, viral DNA extraction and amplification systems will become more rapid, more sensitive, and more automated.

scholarly journals Genotyping of 27 Human Papillomavirus Types by Using L1 Consensus PCR Products by a Single-Hybridization, Reverse Line Blot Detection Method

1998 ◽  
Vol 36 (10) ◽  
pp. 3020-3027 ◽  
Author(s):  
P. E. Gravitt ◽  
C. L. Peyton ◽  
R. J. Apple ◽  
C. M. Wheeler
Keyword(s):  
Human Papillomavirus ◽  
Detection System ◽  
Sampling Error ◽  
Blot Hybridization ◽  
High Sensitivity ◽  
Oligonucleotide Probes ◽  
Dot Blot ◽  
Consensus Primer ◽  
Hpv Dna ◽  
Silver Spring

Amplification of human papillomavirus (HPV) DNA by L1 consensus primer systems (e.g., MY09/11 or GP5+/6+) can detect as few as 10 to 100 molecules of HPV targets from a genital sample. However, genotype determination by dot blot hybridization is laborious and requires at least 27 separate hybridizations for substantive HPV-type discrimination. A reverse blot method was developed which employs a biotin-labeled PCR product hybridized to an array of immobilized oligonucleotide probes. By the reverse blot strip analysis, genotype discrimination of multiple HPV types can be accomplished in a single hybridization and wash cycle. Twenty-seven HPV probe mixes, two control probe concentrations, and a single reference line were immobilized to 75- by 6-mm nylon strips. Each individual probe line contained a mixture of two bovine serum albumin-conjugated oligonucleotide probes specific to a unique HPV genotype. The genotype spectrum discriminated on this strip includes the high-risk, or cancer-associated, HPV genotypes 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 55, 56, 58, 59, 68 (ME180), MM4 (W13B), MM7 (P291), and MM9 (P238A) and the low-risk, or non-cancer-associated, genotypes 6, 11, 40, 42, 53, 54, 57, 66, and MM8 (P155). In addition, two concentrations of β-globin probes allowed for assessment of individual specimen adequacy following amplification. We have evaluated the performance of the strip method relative to that of a previously reported dot blot format (H. M. Bauer et al., p. 132–152, in C. S. Herrington and J. O. D. McGee (ed.), Diagnostic Molecular Pathology: a Practical Approach, (1992), by testing 328 cervical swab samples collected in Digene specimen transport medium (Digene Diagnostics, Silver Spring, Md.). We show excellent agreement between the two detection formats, with 92% concordance for HPV positivity (kappa = 0.78, P < 0.001). Nearly all of the discrepant HPV-positive samples resulted from weak signals and can be attributed to sampling error from specimens with low concentrations (<1 copy/μl) of HPV DNA. The primary advantage of the strip-based detection system is the ability to rapidly genotype HPVs present in genital samples with high sensitivity and specificity, minimizing the likelihood of misclassification.

Human Papillomavirus DNA in Anal CarcinomasComparison of In Situ and Dot Blot Hybridization

1991 ◽  
Vol 96 (3) ◽  
pp. 318-325 ◽  
Author(s):  
MÁire A. Duggan ◽  
Valerie F. Boras ◽  
Masafumi Inoue ◽  
S. Elizabeth McGregor
Keyword(s):  
Human Papillomavirus ◽  
Blot Hybridization ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Papillomavirus Dna

Human papillomavirus in cervix carcinoma and condylomata acuminate-identification of HPV-DNA by improved dot-blot-hybridization

1992 ◽  
Vol 17 (6) ◽  
pp. 392-396 ◽  
Author(s):  
R.M. KNOBLER ◽  
S. SCHNEIDER ◽  
B. RADLWIMMER ◽  
W. BODEMER ◽  
W. GEBHART ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Blot Hybridization ◽  
Cervix Carcinoma ◽  
Dot Blot ◽  
Hpv Dna ◽  
Dot Blot Hybridization

Human Papillomavirus (HPV) Cervical Lesions: Results from 300 Italian Women Studied with Dna Hybridization Techniques and Morphology

1988 ◽  
Vol 74 (6) ◽  
pp. 745-749 ◽  
Author(s):  
Fiorella Nuzzo ◽  
Vittorio Tison ◽  
Antonella Castagnoli ◽  
Maurizio Tiboni ◽  
Ethel-Michele De Villiers ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Human Papillomavirus ◽  
Southern Blot ◽  
Dna Hybridization ◽  
Blot Hybridization ◽  
Hpv Infection ◽  
Cervical Lesions ◽  
Hpv Dna ◽  
Cervical Infection

Human papillomavirus cervical infection was investigated in a series of 300 unselected women by comparing morphological diagnoses (cytology and histology) with results of DNA hybridization techniques (filter in situ hybridization of DNA from exfoliated cervical cells and Southern blot analysis of HPV-DNA in cervical biopsy specimens). The prevalence of HPV cervical infection diagnosed by PAP smears was 11.6 %. Despite disadvantages, filter in situ hybridization was confirmed to be particularly useful for screening purposes to detect HPV in cervical scrapings. In 3 cases it was the only applicable method for diagnosing « high-risk » HPV infection. Southern blot hybridization of tissue DNA with HPV 16-DNA revealed the presence of this virus in 8 cases, and HPV 31-DNA and HPV 42-DNA in 1 case each.

scholarly journals Persistence of Human Papillomavirus DNA in Cervical Lesions After Treatment With Diathermic Large Loop Excision

1998 ◽  
Vol 6 (5) ◽  
pp. 214-219 ◽  
Author(s):  
A. L. Distéfano ◽  
M. A. Picconi ◽  
L. V. Alonio ◽  
D. Dalbert ◽  
J. Mural ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Blot Hybridization ◽  
Simultaneous Detection ◽  
Cervical Lesions ◽  
Dot Blot ◽  
Hpv Dna ◽  
Viral Type ◽  
Large Loop ◽  
Single Stranded Conformational Polymorphism

Objective:The aim of this study was to identify human papillomavirus (HPV) in cervical intraepithelial neoplasia (CIN) lesions and to evaluate the persistence of viral DNA after diathermic large loop excision (DLLE) treatment.Study Design:Biopsies from 36 patients with low- and high-grade CIN lesions were studied before and after DLLE treatment looking for HPV sequences. DNA was extracted to perform a radioactive polymerase chain reaction (PCR) using GP 5,6 generic primers. PCR products were analyzed by the single-stranded conformational polymorphism (SSCP) which is a simultaneous detection and typing method. Dot-blot hybridization with generic and type-specific biotinylated oligonucleotide probes was applied in some cases.Results:HPV DNA was found in all pretreatment samples, and the viral type was identified in 80% of them, HPV 16 being the most prevalent. The viral type coincided with that detected in the first biopsy in all except one case. Seventy five percent of the patients (27 cases) were negative for CIN at follow up, but 50% of them remained HPV DNA positive.Conclusion:DLLE treatment was effective in removing the CIN lesion but not the HPV. This fact points out the need to asses the presence of HPV in DNA during the follow-up, since viral persistence has been considered a high risk factor for recurrence and/or malignant transformation.

A combined ultrastructural and gene molecular research for the relationship between human uterine cervical carcinoma and human papillomavirus

1990 ◽  
Vol 48 (3) ◽  
pp. 204-205
Author(s):  
Kun Lee ◽  
Jingyi Si ◽  
Ricai Han ◽  
Wei Zhang ◽  
Bingbing Tan ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Intraepithelial Neoplasia ◽  
Hpv Infection ◽  
Dot Blot ◽  
Homologous Sequences ◽  
Normal Cervical Epithelium ◽  
The Relationship ◽  
Chronic Cervicitis

There are more supports for the view that human papillomavirus (HPV) infection might be an etiological factor in the development of cervical cancer when the association of persistent condylomata is considered. Biopsies from 318 cases with squamous cell carcinoma of uterine cervix, 48 with cervical and vulvar condylomata, 14 with cervical intraepithelial neoplasia (CIN), 34 with chronic cervicitis and 24 normal cervical epithelium were collected from 5 geographic regions of China with different cervical cancer mortalities. All specimens were prepared for Dot blot, Southern blot and in situ DNA-DNA hybridizations by using HPV-11, 16, 18 DNA labelled with 32P and 3H as probes to detect viral homologous sequences in samples. Among them, 32 cases with cervical cancer, 27 with condyloma and 10 normal cervical epitheliums were randomly chosen for comparative EM observation. The results showed that: 1), 192 out of 318 (60.4%) cases of cervical cancer were positive for HPV-16 DNA probe (Table I)

High Sensitivity of PCR in Situ Hybridization for the Detection of Human Papillomavirus Infection in Uterine Cervical Neoplasias

2001 ◽  
Vol 82 (2) ◽  
pp. 350-354 ◽  
Author(s):  
Yuhong Xiao ◽  
Shigemi Sato ◽  
Takaaki Oguchi ◽  
Kaori Kudo ◽  
Yoshihito Yokoyama ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Human Papillomavirus ◽  
High Sensitivity ◽  
Human Papillomavirus Infection ◽  
Papillomavirus Infection ◽  
Pcr In Situ

Prevalence of HPV in a Melbourne Female STD Population: Comparison of RNA and DNA Probes in Detecting HPV by Dot Blot Hybridization

1993 ◽  
Vol 4 (3) ◽  
pp. 159-164 ◽  
Author(s):  
A J Borg ◽  
G Medley ◽  
S M Garland
Keyword(s):  
Past History ◽  
Blot Hybridization ◽  
Condylomata Acuminata ◽  
Dot Blot ◽  
Hpv Dna ◽  
Sexually Transmitted ◽  
Dot Blot Hybridization ◽  
History Of ◽  
Cytological Features ◽  
Dna Types

A total of 377 women, consecutively selected as first attenders to a sexually transmitted diseases clinic in Melbourne, Australia, were examined for overt Condylomata acuminata and were screened for genital HPV DNA types 6, 11, 16, 18, 31, 33 and (35) using 2 dot blot hybridization methods. Overall, there was a 90% positivity correlation between the 2 methods with HPV DNA being detected in 12% of ectocervical samples. Overt warts were found in 15% of the women and HPV DNA was detected at the cervix in 35% with cytology predicting HPV with or without dysplasia in 27%. Thirteen percent had a past history of warts but none on examination and HPV DNA was evident in 16% while 18% had cytological features of HPV. Those with no warts evident and no past history of warts had both HPV DNA and cytological features of HPV in 7%.

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