Previous Article | Next Article 
Journal of Clinical Microbiology, June 1999, p. 1852-1857, Vol. 37, No. 6
Départements de
Microbiologie-Immunologie et de Pédiatrie,
Received 26 October 1998/Returned for modification 7 December
1998/Accepted 15 March 1999
The line blot assay, a gene amplification method that combines PCR
with nonisotopic detection of amplified DNA, was evaluated for its
ability to detect human papillomavirus (HPV) DNA in genital specimens.
Processed samples were amplified with biotin-labeled primers for HPV
detection (primers MY09, MY11, and HMB01) and for Human papillomavirus (HPV) is now
considered a causative agent of carcinoma of the uterine cervix
(36). Genital HPVs are classified into high-risk types that
are associated with high-grade squamous intraepithelial lesions and
cervical cancer and into low-risk types that are associated with
low-grade squamous intraepithelial lesions (15, 36). HPV DNA
is detected in more than 90% of patients with invasive cancer of the
uterine cervix (4). Cohort studies have demonstrated that
the presence of persistent infection with high-risk HPV types is
predictive of cervical intraepithelial neoplasia and progression to
higher grades of cervical disease (19, 25). Studies of the
natural history of HPV infection, the determinants of persistent HPV
infection, and the prospective impact of a diagnosis of HPV infection
on cervical lesion screening and management are still needed to better
define strategies aimed at preventing and treating precancerous and
cancerous cervical lesions. To reach these objectives, a reliable HPV
detection method that is sensitive and specific and that can be applied
to large numbers of samples from cohort studies is required.
The presence of HPV in clinical specimens is established by nucleic
acid hybridization tests. In order to increase the sensitivity of
detection of HPV DNA, signal amplification and gene amplification methods have been developed (9). The PCR is the most
sensitive method for the detection of HPV DNA sequences in clinical
specimens (5, 16, 31). Since more than 30 HPV types infect
the genital tract, the use of type-specific PCR assays is impractical
for epidemiological studies (1, 12).
The MY09-MY11 consensus primer set targets conserved sequences in the
L1 gene and can amplify a wide spectrum of genital HPV types (3,
4, 20, 24, 28, 30, 33). Amplification products are usually typed
by filter-based assays with type-specific oligonucleotide probes linked
to nonisotopic or isotopic labels (2). Nonisotopic detection
of amplified products facilitates the use of this consensus test for
large-scale testing (2, 10, 26). However, genotype
determination still necessitates several hybridization reactions for
typing, increasing the technical time for analysis of samples.
Consequently, a one-step hybridization procedure would be desirable for
the facilitation of HPV typing.
We report here on an evaluation of the line blot assay (17),
a novel strip-based reverse hybridization test, for the detection of
HPV DNA amplified with MY09-MY11. This nonisotopic assay was evaluated
with clinical specimens obtained in two prospective studies and was
compared to a standard consensus PCR test in which PCR-amplified
products were detected with radiolabeled type-specific probes. Our aims
were to determine the sensitivity and specificity of the line blot test
for detection of the presence of HPV DNA in clinical samples as well as
its reliability for the genotyping of the HPV isolates in HPV-positive samples.
Cell lines and clinical specimens.
The cervical carcinoma
cell line HeLa (which contains 40 copies of HPV type 18 [HPV-18] DNA
per cell) was obtained from the American Type Culture Collection
(Rockville, Md.) and was maintained in Eagle's minimum essential
medium supplemented with 10% fetal calf serum. Two hundred fifty-five
genital specimens were collected from 255 women enrolled in two
different cohort studies investigating the determinants of persistent
HPV infection. One hundred sixty-two cervicovaginal lavage specimens
were from The Canadian Women's HIV study (8, 18). This
study evaluates the relationship between genital HPV infection and
cervical disease progression in relation to human immunodeficiency
virus (HIV)-induced immune deficiency. The cervicovaginal lavage
specimens were selected on the basis of initial results obtained with
MY09-MY11 amplification reactions and detection with isotopic
type-specific probes. This selection ensured the inclusion of all HPV
types detected in the standard consensus PCR test and ensured the
inclusion of specimens containing multiple HPV types. Ninety-three
consecutive cervical brushings were from an ongoing study on the
determinants of HPV persistence in young adult women attending McGill
University (29). All samples were tested by the line blot
assay and the standard consensus PCR test without knowledge of previous
results or the patients' clinical status. Consent was obtained from
each participant. Both projects had the approval of the ethics
committees of the institutions involved.
Processing of clinical samples.
Cervicovaginal lavage
specimens were obtained with 10 ml of phosphate-buffered saline (pH
7.4) by standard procedures (35). The cells were then
centrifuged in an IEC Centra-8R centrifuge at 2,500 rpm for 10 min at
4°C, resuspended in 500 µl of 10 mM Tris-HCl (pH 8.2), and stored
frozen at Standard consensus PCR test.
HPV DNA was amplified under
standard conditions with the MY09, MY11, and HMB01 consensus HPV
primers as described previously (8, 11, 20). Amplifications
of HPV DNA and Line blot assay.
Lysates were amplified by the consensus L1
PCR protocol described above, with the following modifications made to
the amplification mixture (17): the use of 6 mM
MgCl2, 7.5 U of AmpliTaq Gold DNA polymerase, 600 µM
dUTP, and 200 µM each dATP, dCTP, and dGTP, and biotin-labeled
primers (MY09, MY11, HMB01, PC04, and GH20). A rapid amplification
profile was used in a TC 9600 thermal cycler (Perkin-Elmer Cetus):
activation of AmpliTaq Gold at 95°C for 9 min; denaturation at 95°C
for 20 s, annealing at 55°C for 30 s, and extension at
72°C for 30 s for 40 cycles; and terminal extension at 72°C
for 5 min (17).
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Nonisotopic Detection and Typing of Human
Papillomavirus DNA in Genital Samples by the Line Blot Assay
ABSTRACT
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
-globin detection
(primers PC03 and PC04). Amplified DNA products were hybridized by a
reverse blot method with oligonucleotide probe mixtures fixed on a
strip that allowed the identification of 27 HPV genotypes. The line
blot assay was compared to a standard consensus PCR test in which HPV
amplicons were detected with radiolabeled probes in a dot blot assay.
Two hundred fifty-five cervicovaginal lavage specimens and cervical
scrapings were tested in parallel by both PCR tests. The line blot
assay consistently detected 25 copies of HPV type 18 per run. The
overall positivity for the DNA of HPV types detectable by both methods
was 37.7% (96 of 255 samples) by the line blot assay, whereas it was
43.5% (111 of 255 samples) by the standard consensus PCR assay. The
sensitivity and specificity of the line blot assay reached 84.7% (94 of 111 samples) and 98.6% (142 of 144 samples), respectively. The
agreement for HPV typing between the two PCR assays reached 83.9% (214 of 255 samples). Of the 37 samples with discrepant results, 33 (89%) were resolved by avoiding coamplification of -globin and modifying the amplification parameters. With these modifications, the line blot
assay compared favorably to an assay that used radiolabeled probes. Its
convenience allows the faster analysis of samples for large-scale
epidemiological studies. Also, the increased probe spectrum in this
single hybridization assay permits more complete type discrimination.
INTRODUCTION
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
MATERIALS AND METHODS
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
70°C until they were processed (7). The cell
suspensions were thawed, lysed by the addition of Tween 20 and Nonidet
P-40 (each at a final concentration of 0.4% [vol/vol]), and digested
with 250 µg of proteinase K per ml for 2 h at 45°C. Cell
lysates were boiled for 10 min and stored at 70°C until they were tested.
70°C until they were tested. Five microliters of
processed sample was tested in each PCR assay. All samples had tested
positive for -globin with primers PC04 and GH20 (3, 8).
-globin DNA were done in separate reactions. The
amplification mixture contained 6.5 mM MgCl2, 50 mM KCl,
2.5 U of Taq DNA polymerase (AmpliTaq; Roche Molecular
Diagnostics, Mississauga, Ontario, Canada), 200 µM each dATP, dCTP,
dGTP, and dTTP, and 50 pmol of each primer. Negative and weakly
positive (25 HPV-18 DNA copies) controls were included to monitor
contamination and the overall endpoint sensitivity of each PCR run.
Amplifications were performed in a 480 Thermocycler (Perkin-Elmer
Cetus, Montréal, Quebec, Canada) for 40 cycles with the following
cycling parameters: 95°C for 1 min, 55°C for 1 min, and 72°C for
1 min. The amplified products were spotted onto nylon membranes and
were reacted under stringent conditions with 32P-labeled
oligonucleotide probes for types 6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, and 58 (2, 3, 20). The measures used to avoid
false-positive reactions due to contamination have been described
elsewhere (8).
-globin
probes. The probe mixtures for the following 27 HPV genotypes were
fixed on distinct lines on each strip: types 6, 11, 16, 18, 26, 31, 33, 35, 39, 40, 42, 45, 51, 52, 53, 54, 55, 56, 57, 58, 59, 66, 68, MM4,
MM7, MM8, and MM9. The tray was incubated in a shaking water bath at
53°C for 30 min. The hybridization solution was aspirated from each
well, and 3 ml of washing solution containing 1× SSPE and 0.1% sodium
dodecyl sulfate was added at room temperature and was aspirated. Again,
3 ml of washing solution was added to each well and the plate was
incubated at 53°C for 15 min. The washing buffer was aspirated, and 3 ml of AMPLICOR streptavidin-horseradish peroxidase conjugate was added.
The tray was shaken gently for 30 min at room temperature. The
conjugate was aspirated and 3 ml of washing buffer was added. The trays
were shaken for 10 min on a platform shaker. This step was repeated
once. After aspiration of the washing buffer, 3 ml of citrate buffer
was added to each well and was aspirated. The substrate was prepared by
mixing 0.01% H2O2 and 0.1% ProClin in a 0.1 M
citrate solution with 0.1% tetramethylbenzidine in dimethylformamide.
Three milliliters of substrate was added to each well. The trays were
shaken at 70 rpm for 5 min at room temperature. The substrate was
removed, the strips were rinsed with distilled water and stored in
citrate buffer, and the results were read within 30 min.
-globin
primers to the amplification reaction mixture. These lysates were also
tested by the line blot assay by using the ultrasensitive amplification
cycling profile (17) (activation of AmpliTaq Gold at 95°C
for 9 min; denaturation at 95°C for 60 s, annealing at 55°C
for 60 s, and extension at 72°C for 60 s for 40 cycles; and
terminal extension at 72°C for 5 min).
Statistical method. The crude percent agreement between both detection methods was the percentage of paired samples with identical results. The agreements for overall positivity (HPV DNA positive), for positivity for high-risk HPV types as a group (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, and 58), and for positivity for each type were calculated. The unweighted kappa statistic was calculated to adjust for chance agreement between HPV detection methods (14). In general, a kappa value above 0.75 represents excellent agreement beyond chance. The sensitivity and specificity of the line blot assay were calculated by considering the standard consensus PCR test as the "gold standard." The mean number of types detected per sample by each PCR test was compared by the Mann-Whitney rank sum test, since the distribution of types per sample was not normally distributed. Chi-square analysis was performed to compare proportions.
| |
RESULTS |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
HPV DNA detection. HPV DNA was detected in 47.5% (121 of 255) and 43.5% (111 of 255) of samples by using the line blot assay and the standard consensus PCR test, respectively. However, only 14 genotypes (types 6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, and 58) were identified by both assays. The following analyses were restricted to these 14 types detectable by the standard consensus and the line blot assays. Specimens that tested positive by the line blot assay for an HPV type not included in the latter 14 types were considered negative in the comparisons.
First, the line blot and the standard consensus PCR assays were compared for their abilities to detect the presence of HPV DNA of the 14 detectable types enumerated above in 255 genital specimens. HPV DNA was detected in 96 (37.7%) and 111 (43.5%) of the 255 samples by the line blot assay and the standard consensus PCR test, respectively. Identical results were obtained by both tests for 236 (92.6%) of 255 samples (94 HPV-positive samples and 142 HPV-negative samples) for a very good agreement for the detection of the presence of HPV DNA in genital samples (kappa statistic of 0.82). Of the 113 samples positive for HPV DNA by at least one PCR method, HPV DNA was detected by one method only in 19 samples. In two of these samples, infections with single HPV types (once each with HPV-39 and HPV-53) were detected only by the line blot assay. The testing of biotin-labeled amplified products from these two samples with isotopic probes confirmed the presence of these HPV types. Repeat testing of the two samples by the line blot assay also confirmed the presence of HPV DNA, but the standard consensus PCR test remained negative. In the other 17 samples, the presence of HPV DNA from 16 infections caused by single HPV types and from 1 infection caused by several HPV types was detected only by the standard consensus PCR test. Biotin-labeled PCR products from line blot assays for these samples were tested by a dot blot assay with radiolabeled probes and scored negative for HPV. This last experiment confirmed that these samples were falsely negative by the line blot assay because of differences in the amplification process between the two PCR assays and not because of a lack of sensitivity of the strip hybridization assay. When the standard consensus PCR was considered to be the gold standard, the sensitivity and the specificity of the line blot assay for detection of the presence of HPV DNA reached 84.7% (94 of 111) and 98.6% (142 of 144), respectively.HPV DNA type identification.
The distributions of the 27 types
detected by the line blot assay and the 14 types detected by the
standard consensus PCR assay are presented in Table
1. When only the 14 HPV types detectable by both assays were considered, 46 samples (40.7% of the 113 HPV-positive samples) contained multiple HPV types by at least one PCR
assay, while 32 contained multiple types by both assays.
|
HPV typing results by line blot assay by avoiding
coamplification.
HPV and -globin sequences were coamplified
only by the line blot assay. Since in our experience coamplification
reduces the level of sensitivity for HPV detection with consensus L1
primers (6), we avoided -globin coamplification in the
standard consensus PCR assay. When coamplification of -globin and
HPV DNA was done by the line blot assay, 10 copies of HPV-18 scored
negative in two of five PCR runs (data not shown). The same positive
control scored positive in five of five PCR runs when primers for
-globin were not added to the master mixture (data not shown). In 37 samples, the line blot assay detected fewer types than the standard
consensus PCR test. Retesting of these samples by the line blot and the standard consensus PCR assays yielded the same results as those obtained in the first amplification run (data not shown). The latter 37 samples were again amplified by the line blot assay but the
-globin primers were not added to the amplification mixture.
-globin: the results remained
identical to those obtained in the first amplification run (data not
shown). When only the presence or absence of HPV DNA is considered, 12 of the 17 specimens initially negative by the line blot assay but
positive by the standard consensus PCR test were positive by the line
blot assay, for a sensitivity of the line blot assay of 95.5% (106 of
111 samples). When typing results are considered, 21 of the 37 discrepancies (56.8%) were resolved. Of the 21 samples with resolved
discrepancies, 9 contained more than one HPV type by the standard
consensus PCR assay (the types resolved included type 56 for three
women, type 6 for one woman, type 31 for one woman, type 33 for one
woman, type 51 for one woman, type 52 for one woman, and type 58 for
one woman) and 12 contained one type by the standard consensus PCR
assay (type 56 for three women, type 58 for three women, type 6 for one
woman, type 16 for one woman, type 33 for one woman, type 35 for one woman, type 52 for one woman, and type 53 for one woman). Results from
two additional samples that contained multiple types were partly
resolved. In the latter samples, types 31 and 39 were detected by the
line blot assay without coamplification but still remained falsely
negative for types 51 and 56. The results for 14 samples remained
unresolved, including 9 samples with more than one type (types
undetected by the modified line blot assay included HPV-56 for 3 samples and HPV-51, -33, -39, -35, -58, -52, and -53 for one sample
each) and 5 samples with one type by the standard consensus PCR assay
(the types undetected by the modified line blot assay included HPV-52
for 2 samples, HPV-35 for 2 samples, and HPV-18 for 1 sample). The mean
number of types per sample containing multiple types that remained
unresolved was similar to that for samples containing multiple types
that were resolved (2.8 ± 1.4 types versus 2.6 ± 0.6 types;
P = 0.715).
Next, the impact of avoiding coamplification was investigated by using
the ultrasensitive amplification profile for these 37 samples
(17). In this profile, each step of the 40 cycles of
amplification is extended to 1 min instead of 20 or 30 s.
Discrepancies were resolved for 33 (89.2%) samples and were partly
resolved for 1 sample (Table 1). By combining these new results
obtained by the line blot assay without -globin primers with initial
results obtained by the line blot assay for samples with concordant
results, the sensitivity of the line blot assay without coamplification for the detection of HPV types reached 96.3% (105 of 109 samples; 95%
confidence interval, 92.6 to 100.0%).
| |
DISCUSSION |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
This report describes the evaluation of a practical
nonisotopic method for the typing of HPV L1 DNA amplified by PCR from exfoliated cells obtained by cervicovaginal lavage or from cervical scraping. The method involves a reverse hybridization reaction of
biotin-labeled amplified DNA with oligonucleotide probes fixed on a
strip. We have compared this novel assay to a standardized test that
served as a gold standard. Both assays used the same primer pair that
has demonstrated good levels of sensitivity and reproducibility in
other studies (27). Several differences are found between
the line blot assay and the standard consensus PCR test.
Coamplification of HPV and -globin DNA was accomplished only by the
line blot assay. The line blot assay uses a novel thermostable DNA
polymerase that is activated by high temperatures and that does not
require hot start. In the line blot assay, biotin-labeled PCR products
are reacted to probes fixed onto lines on a strip. The standard
consensus PCR test detected with radiolabeled probes amplicons fixed on
nylon membranes. The line blot assay can be completed in a short time
because amplified DNA is hybridized with all relevant probes in one
reaction instead of several reactions.
There was a good agreement between these two assays. The agreement
exceeded 90% for HPV DNA detection, detection of high-risk HPV types
as a group, and typing of HPV when coamplification was not done. The
line blot assay was not as sensitive as the standard consensus PCR
assay, especially with samples with more than one HPV type. Samples
with discordant results between the two PCR tests were more likely to
be infected with multiple types. An important proportion of these
discrepancies were found with HPV-56 or -58 infection and rarely
affected frequently encountered high-risk types such as HPV-16.
Avoidance of coamplification and the use of the ultrasensitive
amplification profile (17) resolved most (89%) of the
discrepancies between the two PCR tests. Although faster, the rapid
amplification profile should not be used for the sensitive detection of
HPV DNA. In the first evaluation of the line blot assay
(17), the rapid amplification profile was also less
sensitive that the ultrasensitive profile. Previous work with the line
blot assay did not report a loss of sensitivity due to coamplification
(17). This variability in the performance of the line blot
assay could be in part related to the strategy of synthesis of
biotin-labeled primers: batch synthesis of biotin-labeled primers was
not controlled to ensure an equivalent representation and synthesis of
all degenerate primers. Underrepresentation of some degenerate primers
could lead to inefficient amplification for some types. The use of a
rapid amplification profile could also explain the loss in sensitivity
with coamplification. We did not evaluate coamplification of HPV and
-globin DNA using the ultrasensitive profile because limited amounts
of sample remained. The line blot assay had an excellent specificity,
considering that biotin-labeled PCR products from line blot
assay-positive and standard consensus PCR-negative samples reacted with
radiolabeled probes in a dot blot assay.
The performance of nonisotopic hybridization assays is often comparable to that of methods that use radiolabeled probes for detection of HPV DNA amplified products (2, 10, 21-23, 32). The use of nonisotopic probes in filter-based hybridization assays has been described with consensus PCR tests, but successive tests with individual probes for each type were still required (2, 27, 37). Manipulation of strips in the line blot assay is more convenient than the handling of filters. Some nonisotopic tests (10, 21, 23, 32) have the advantage of using 96-well microtiter plates which are easier to manipulate and which are more convenient than filter-based assays. However, most require the testing of samples with multiple individual probes and could not detect as many types as the number described here for the line blot assay with MY09-MY11 PCR products.
A reverse hybridization assay has also been described for HPV detection (34). Compared to the line blot assay, complete HPV genomic probes were fixed on membranes instead of type-specific oligonucleotide probes. Because a limited number of types were evaluated and because of the results of previous work demonstrating cross-hybridization when long probes are used to type MY09-MY11 amplicons (10, 13), cross-hybridization could still be encountered in that system.
A substantial number of samples contained HPV types that have been recently described, such as types 66, MM7, and MM8. This could result from the selection of HPV-positive samples. However, these results underscore the importance of keeping assays current as new types are discovered.
In conclusion, the line blot assay compared favorably to a standard isotopic method for detection of PCR products with clinical specimens. It was nearly as sensitive as the standard radioisotopic assay even with samples infected with multiple HPV types, especially if coamplification was avoided and if the ultrasensitive amplification profile was used. This convenient format allows one to test one sample for 27 types in one reaction.
| |
ACKNOWLEDGMENTS |
|---|
We thank Diane Gaudreault and Diane Bronsard for processing the genital samples.
This work was supported in part by Roche Molecular Systems, which supplied reagents for the line blot assay.
The Medical Research Council of Canada and Health and Welfare Canada support The Canadian Women's HIV study. The Medical Research Council of Canada supports the HPV persistence study in university students. F.C. is a clinical research scholar supported by FRSQ, and E.F. is a research scholar supported by FRSQ.
| |
FOOTNOTES |
|---|
* Corresponding author. Mailing address: Département de Microbiologie et Infectiologie, Centre Hospitalier de l'Université de Montréal, Campus Notre-Dame, 1560 Sherbrooke est, Montréal, Québec H2L 4M1, Canada. Phone: 514-281-6000, ext. 5162. Fax: 514-896-4607. E-mail: coutleef{at}sympatico.ca.
Present address: Department of Epidemiology, Johns Hopkins
University School of Hygiene and Public Health, Baltimore, MD 21205.
The Canadian Women's HIV study group includes the following
investigators: Catherine Hankins, Normand Lapointe, John Gill, Barbara
Romanowski, Stephen Shafran, Rob Grimshaw, David Haase, Wally Schlech,
Stephen Landis, John Sellors, Fiona Smaill, Francois Beaudoin, Marc
Boucher, Ngoc Bui, Michel Chateauvert, Manon Coté, François
Coutlée, Douglas Dalton, Gretty Deutsch, Julian Falutz, Diane
Francoeur, Lisa Hallman, Lina Karayan, Louise Labrecque, Richard
Lalonde, Christiane Lavoie, Catherine Lounsbury, John Macleod, Nicole
Marceau, Grégoire Noel, Grégoire Piché, Jean-Pierre Routy, Pierre Simard, Christina Smeja, Graham Smith, Pierre-Paul Tellier, Emil Toma, Garry Garber, Garry Victor, Louise Coté, Édith Guilbert, Michel Morissette, Hélène Senay,
Sylvie Trottier, Phil Berger, Lisa Friedland, Donna Keystone, Joan
Murphy, Anne Phillips, Marion Powell, Anita Rachlis, Pat Rockman,
Irving Salit, Cheryl Wagner, Sharon Walmsey, Kurt Williams, Ian Bowmer,
Rory Windrim, Roger Sandre, Penny Ballem, David Burdge, Brian Conway, Marianne Harris, Deborah Money, Julio Montaner, and Janice Veenhuizen.
| |
REFERENCES |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
| 1. |
Alani, R. M., and K. Munger.
1998.
Human papillomaviruses and associated malignancies.
J. Clin. Oncol.
16:330-337 |
| 2. | Bauer, H. M., C. E. Greer, and M. M. Manos. 1992. Determination of genital human papillomavirus infection by consensus polymerase chain reaction amplification, p. 131-152. In C. S. Herrington, and J. O. D. McGee (ed.), Diagnostic molecular pathology, a practical approach. IRL Press, Oxford, United Kingdom. |
| 3. |
Bauer, H. M.,
Y. Ting,
C. E. Greer,
J. C. Chambers,
C. J. Tashiro,
J. Chimera,
A. Reingold, and M. M. Manos.
1991.
Genital human papillomavirus infection in female university students as determined by a PCR-based method.
JAMA
265:472-477 |
| 4. |
Bosch, F. X.,
M. M. Manos,
N. Muñoz,
M. Sherman,
A. M. Jansen,
J. Peto,
M. H. Schiffman,
V. Moreno,
R. Kurman,
K. V. Shah, and IBSCC Study Group.
1995.
Prevalence of human papillomavirus in cervical cancer: a worldwide perspective.
J. Natl. Cancer Inst.
87:796-802 |
| 5. | Cope, J. U., A. Hildesheim, M. H. Schiffman, M. M. Manos, A. T. Lorincz, R. D. Burk, A. G. Glass, C. Greer, J. Buckland, K. Helgesen, D. R. Scott, M. E. Sherman, R. J. Kurman, and K. L. Liaw. 1997. Comparison of the hybrid capture tube test and PCR for detection of human papillomavirus DNA in cervical specimens. J. Clin. Microbiol. 35:2262-2265[Abstract]. |
| 6. | Coutlée, F. Personal communication. |
| 7. | Coutlée, F., L. Bobo, A. Hawwari, G. Dalabetta, N. E. Hook, K. V. Shah, and R. P. Viscidi. 1992. Detection of HPV-16 in cell lines and cervical lavages specimens by a polymerase chain reaction-enzyme immunoassay assay. J. Med. Virol. 37:22-29[Medline]. |
| 8. | Coutlée, F., C. Hankins, N. Lapointe, and The Canadian Women's HIV Study. 1997. Comparison between vaginal tampon and cervicovaginal lavage specimen collection for detection of human papillomavirus DNA by the polymerase chain reaction. J. Med. Virol. 51:42-47[Medline]. |
| 9. | Coutlée, F., M. H. Mayrand, D. Provencher, and E. Franco. 1997. The future of HPV testing in clinical laboratories and applied virology research. Clin. Diagn. Virol. 8:123-141[Medline]. |
| 10. | Coutlée, F., D. Provencher, and H. Voyer. 1995. Detection of human papillomavirus DNA in cervical lavage specimens by a nonisotopic consensus PCR assay. J. Clin. Microbiol. 33:1973-1978[Abstract]. |
| 11. | Coutlée, F., A. M. Trottier, G. Ghattas, R. Leduc, E. Toma, G. Sanche, I. Rodrigues, B. Turmel, G. Allaire, and P. Ghadirian. 1997. Risk factors for oral human papillomavirus in adults infected and not infected with human immunodeficiency virus. Sex. Transm. Dis. 24:23-31[Medline]. |
| 12. |
de Villiers, E. M.
1989.
Heterogeneity of the human papillomavirus group.
J. Virol.
63:4898-4901 |
| 13. | Digene Diagnostics Inc. 1993. Digene SHARP signal system-HPV probe and primer set. Package insert. Digene Diagnostics Inc., Beltsville, Md. |
| 14. | Fleiss, J. L. 1981. Statistical methods for rates and proportions. John Wiley & Sons, Inc., New York, N.Y. |
| 15. | Franco, E. L. F. 1996. Epidemiology of anogenital warts and cancer, p. 597-623. In C. P. Crum, and A. T. Lorincz (ed.), Human papillomavirus. The W. B. Saunders Co., New York, N.Y. |
| 16. | Gravitt, P., A. Hakenewerth, and J. Stoerker. 1991. A direct comparison of methods proposed for use in widespread screening of human papillomavirus infection. Mol. Cell. Probes 5:65-72[Medline]. |
| 17. |
Gravitt, P.,
C. L. Peyton,
R. J. Apple, and C. Wheeler.
1998.
Genotyping of 27 human papillomavirus types by using L1 consensus PCR products by single-hybridization, reverse line blot detection method.
J. Clin. Microbiol.
36:3020-3027 |
| 18. | Hankins, C., F. Coutlee, N. Lapointe, P. Simard, T. Tran, J. Samson, L. Hum, and The Canadian Women's HIV Study Group. Risk factors associated with the human papillomavirus infection in women living with HIV. Can. Med. Assoc. J., in press. |
| 19. | Herrington, C. S., M. F. Evans, N. F. Hallam, F. M. Charnock, W. Gray, and D. McGee. 1995. Human papillomavirus status in the prediction of high-grade cervical intraepithelial neoplasia in patients with persistent low-grade cervical cytological abnormalities. Br. J. Cancer 71:206-209[Medline]. |
| 20. | Hildesheim, A., M. H. Schiffman, P. E. Gravitt, A. G. Glass, C. E. Greer, T. Zhang, D. R. Scott, B. B. Rush, P. Lawler, M. E. Sherman, R. J. Kurman, and M. M. Manos. 1994. Persistence of type-specific human papillomavirus infection among cytologically normal women. J. Infect. Dis. 169:235-240[Medline]. |
| 21. | Jacobs, M. V., P. J. F. Snidjers, A. J. C. van den Brule, T. J. M. Helmerhorst, C. Meijer, and J. Walboomers. 1997. A general primer GP5+/GP6+-mediated PCR-enzyme immunoassay method for rapid detection of 14 high-risk and 6 low-risk human papillomavirus genotypes in cervical scrapings. J. Clin. Microbiol. 35:791-795[Abstract]. |
| 22. | Jacobs, M. V., A. J. C. Vandenbrule, P. J. F. Snijders, T. J. M. Helmerhorst, C. J. L. M. Meijer, and J. M. M. Walboomers. 1996. A non-radioactive PCR enzyme-immunoassay enables a rapid identification of HPV16 and 18 in cervical scrapes after GP5+/6+ PCR. J. Med. Virol. 49:223-229[Medline]. |
| 23. | Lazar, J. G., A. J. Tumulty, H. Salim, and S. S. Challberg. 1993. Sensitive and specific detection of PCR products: application of the Digene SHARP signal system, abstr. C-218, p. 484. In Abstracts of the 93rd General Meeting of the American Society for Microbiology 1993. American Society for Microbiology, Washington, D.C. |
| 24. |
Ley, C.,
H. M. Bauer,
A. Reingold,
M. H. Schiffman,
J. C. Chambers,
C. J. Tashiro, and M. M. Manos.
1991.
Determinants of genital human papillomavirus infection in young women.
J. Natl. Cancer Inst.
83:997-1003 |
| 25. | Londesborough, P., L. Ho, G. Terry, J. Cuzick, C. Wheeler, and A. Singer. 1996. Human papillomavirus genotype as a predictor of persistence and development of high-grade lesions in women with minor cervical abnormalities. Int. J. Cancer 69:364-368[Medline]. |
| 26. | Manos, M. M., Y. Ting, D. K. Wright, A. J. Lewis, T. Broker, and S. M. Wolinski. 1989. Use of the polymerase chain reaction amplification for the detection of genital human papillomaviruses. Cancer Cells 7:209-215. |
| 27. | Qu, W., G. Jiang, Y. Cruz, C. J. Chang, G. Y. F. Ho, R. S. Klein, and R. D. Burk. 1997. PCR detection of human papillomavirus: comparison between MY09/MY11 and GP5+/GP6+ primer systems. J. Clin. Microbiol. 35:1304-1310[Abstract]. |
| 28. |
Resnick, R. M.,
M. T. E. Cornelissen,
D. K. Wright,
G. H. Eichinger,
H. S. Fox,
J. T. Schegget, and M. M. Manos.
1990.
Detection and typing of human papillomavirus in archival cervical cancer specimens by DNA amplification with consensus primers.
J. Natl. Cancer Inst.
82:1477-1484 |
| 29. | Richardson, H., J. Pintos, F. Coutlée, P. Tellier, P. Gravitt, and E. Franco. 1998. Risk factors for persistent cervical human papillomavirus infection in Montreal university students. In Abstracts of the First International Conference on Human Papillomavirus Infections and Cervical Cancer. |
| 30. |
Schiffman, M. H.,
H. M. Bauer,
R. N. Hoover,
A. G. Glass,
D. M. Cadell,
B. B. Rush,
D. R. Scott,
M. E. Sherman,
R. J. Kurman,
S. Wacholder,
C. K. Stanton, and M. M. Manos.
1993.
Epidemiologic evidence showing that human papillomavirus infection causes most cervical intraepithelial neoplasia.
J. Natl. Cancer Inst.
85:958-963 |
| 31. |
Schiffman, M. H.,
H. M. Bauer, and A. T. Lorincz.
1991.
Comparison of Southern blot hybridization and polymerase chain reaction methods for the detection of human papillomavirus DNA.
J. Clin. Microbiol.
29:573-577 |
| 32. | Terry, G., L. Ho, A. Szarewaki, and J. Cuzick. 1995. Semi-automated detection of human papillomavirus DNA of high and low oncogenic potential in cervical smears, in press. Imperial Cancer Research Fund, London, United Kingdom. |
| 33. | Ting, Y., and M. Manos. 1990. Detection and typing of genital human papillomavirus, p. 356-370. In PCR protocols: a guide to methods and applications. Academic Press, New York, N.Y. |
| 34. | Venuti, A., G. Badaracco, and M. L. Marcante. 1995. Detection and typing of human papillomavirus by single hybridization. J. Virol. Methods 51:115-124[Medline]. |
| 35. | Vermund, S. H., M. H. Schiffman, G. L. Goldberg, D. B. Ritter, A. Weltman, and R. D. Burk. 1989. Molecular diagnosis of genital human papillomavirus infection comparison of two methods used to collect exfoliated cervical cells. Am. J. Obstet. Gynecol. 160:304-308[Medline]. |
| 36. | Villa, L. L. 1997. Human papillomaviruses and cervical cancer. Adv. Cancer Res. 71:321-341[Medline]. |
| 37. | Ylitalo, N., T. Bergstrom, and U. Gyllensten. 1995. Detection of genital human papillomavirus by single-tube nested PCR and type-specific oligonucleotide hybridization. J. Clin. Microbiol. 33:1822-1828[Abstract]. |
Journal of Clinical Microbiology, June 1999, p. 1852-1857, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Coutlee, F., Rouleau, D., Ghattas, G., Hankins, C., Vezina, S., Cote, P., Macleod, J., de Pokomandy, A., Money, D., Walmsley, S., Voyer, H., Brassard, P., Franco, E.
(2007). Confirmatory Real-Time PCR Assay for Human Papillomavirus (HPV) Type 52 Infection in Anogenital Specimens Screened for HPV Infection with the Linear Array HPV Genotyping Test. J. Clin. Microbiol.
45: 3821-3823
[Abstract] [Full Text] -
Coutlee, F., Rouleau, D., Petignat, P., Ghattas, G., Kornegay, J. R., Schlag, P., Boyle, S., Hankins, C., Vezina, S., Cote, P., Macleod, J., Voyer, H., Forest, P., Walmsley, S., The Canadian Women's HIV Study Group, , Franco, E.
(2006). Enhanced Detection and Typing of Human Papillomavirus (HPV) DNA in Anogenital Samples with PGMY Primers and the Linear Array HPV Genotyping Test.. J. Clin. Microbiol.
44: 1998-2006
[Abstract] [Full Text] -
Huang, S.-L., Chao, A., Hsueh, S., Chao, F.-Y., Huang, C.-C., Yang, J.-E., Lin, C.-Y., Yan, C.-C., Chou, H.-H., Huang, K.-G., Huang, H.-J., Wu, T.-I, Tseng, M.-J., Qiu, J.-T., Lin, C.-T., Chang, T.-C., Lai, C.-H.
(2006). Comparison between the Hybrid Capture II Test and an SPF1/GP6+ PCR-Based Assay for Detection of Human Papillomavirus DNA in Cervical Swab Samples.. J. Clin. Microbiol.
44: 1733-1739
[Abstract] [Full Text] -
Richardson, H., Kelsall, G., Tellier, P., Voyer, H., Abrahamowicz, M., Ferenczy, A., Coutlee, F., Franco, E. L.
(2003). The Natural History of Type-specific Human Papillomavirus Infections in Female University Students. Cancer Epidemiol. Biomarkers Prev.
12: 485-490
[Abstract] [Full Text] -
Kornegay, J. R., Roger, M., Davies, P. O., Shepard, A. P., Guerrero, N. A., Lloveras, B., Evans, D., Coutlee, F.
(2003). International Proficiency Study of a Consensus L1 PCR Assay for the Detection and Typing of Human Papillomavirus DNA: Evaluation of Accuracy and Intralaboratory and Interlaboratory Agreement. J. Clin. Microbiol.
41: 1080-1086
[Abstract] [Full Text] -
Bosch, F X, Lorincz, A, Munoz, N, Meijer, C J L M, Shah, K V
(2002). The causal relation between human papillomavirus and cervical cancer. J. Clin. Pathol.
55: 244-265
[Abstract] [Full Text] -
van den Brule, A. J. C., Pol, R., Fransen-Daalmeijer, N., Schouls, L. M., Meijer, C. J. L. M., Snijders, P. J. F.
(2002). GP5+/6+ PCR followed by Reverse Line Blot Analysis Enables Rapid and High-Throughput Identification of Human Papillomavirus Genotypes. J. Clin. Microbiol.
40: 779-787
[Abstract] [Full Text] -
Coutlee, F., Gravitt, P., Kornegay, J., Hankins, C., Richardson, H., Lapointe, N., Voyer, H., Franco, E.
(2002). Use of PGMY Primers in L1 Consensus PCR Improves Detection of Human Papillomavirus DNA in Genital Samples. J. Clin. Microbiol.
40: 902-907
[Abstract] [Full Text] -
Kornegay, J. R., Shepard, A. P., Hankins, C., Franco, E., Lapointe, N., Richardson, H., Canadian Women's HIV Study Group, , Coutlee, F.
(2001). 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.
39: 3530-3536
[Abstract] [Full Text] -
Coutlée, F., de Ladurantaye, M., Tremblay, C., Vincelette, J., Labrecque, L., Roger, M.
(2000). An Important Proportion of Genital Samples Submitted for Chlamydia trachomatis Detection by PCR Contain Small Amounts of Cellular DNA as Measured by beta -Globin Gene Amplification. J. Clin. Microbiol.
38: 2512-2515
[Abstract] [Full Text] -
Vernon, S. D., Unger, E. R., Williams, D.
(2000). Comparison of Human Papillomavirus Detection and Typing by Cycle Sequencing, Line Blotting, and Hybrid Capture. J. Clin. Microbiol.
38: 651-655
[Abstract] [Full Text] -
Lacey, J. V. Jr., Brinton, L. A., Abbas, F. M., Barnes, W. A., Gravitt, P. E., Greenberg, M. D., Greene, S. M., Hadjimichael, O. C., McGowan, L., Mortel, R., Schwartz, P. E., Silverberg, S. G., Hildesheim, A.
(1999). Oral Contraceptives as Risk Factors for Cervical Adenocarcinomas and Squamous Cell Carcinomas. Cancer Epidemiol. Biomarkers Prev.
8: 1079-1085
[Abstract] [Full Text]
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»