0095-1137/90/092076-06$02.00/0
Use of AffiProbe HPV
Test Kit for Detection of
Human
Papillomavirus
DNA in Genital
Scrapes
MARJUT RANKI,l* ANTTI W. LEINONEN,' TARJA JALAVA,' PEKKA NIEMINEN,2 VALÉRIA R. X. SOARES,2 JORMAPAAVONEN,2AND ARJAKALLIO1
OrionPharmaceutica/Biotechnology, OrionCorporation,' andDepartment ofObstetrics andGynecology,
University
CentralHospital,2
Helsinki, FinlandReceived13 February1990/Accepted 25 May 1990
The presence of human papillomavirus (HPV) DNA in cervical and vaginal scrapes was analyzed by the AffiProbe HPV test kit (Orion Corp., Orion Pharmaceutica, Helsinki, Finland), which is a 1-day solution hybridizationtestforHPV type6/11, 16,or18.TheAffiProbetestwascomparedwithacommerciallyavailable
dot blot test (ViraPap and ViraType tests; Life Technologies Inc., Gaithersburg, Md.). The study group consistedof 178patientsseeninagynecological outpatientclinic.Altogether,64specimens(36cervical and28 vaginal scrapes) from 49patientswerepositive bythe AffiProbe test.Concurrently collected cervicalscrapes from 174patientswereavailable for thereferencetest,whichyielded27positiveresultsfor HPVtype6/11or
16/18 and25positive resultsfor HPV type31/33135. Agreementasto thepresenceof HPVtype6/11, 16,or18 bythetwotestswasreached in85 %of thespecimens.Eleven cervicalspecimenswerepositivebytheAffiProbe testonly,and nine cervicalspecimenswerepositivebytheViraTypetestonly.Independentevidenceobtained bythepolymerase chainreaction, repeatexamination, orthe concurrentpresenceof HPVDNAinvaginalor
vulvalepithelium supportedtheAffiProbe and theViraTypetestresults for 6 of the11and 6of the9specimens withdiscrepant results, respectively. Thus, the DNAtests had similar sensitivities for HPVtype 6/11, 16,and 18DNAs,but theresultswereobtained within 1day bythe AffiProbetest,whereas results fortheViraPapand ViraType analyses requiredfrom 4 daysto2 weeks.
Human papillomaviruses (HPVs) cause infections ofthe genitaltract, whichcanbesubclinicalormanifestasvisible
orflat condylomas. HPVinfectionsarecommon;theannual incidence inFinland is 8% in young, sexuallyactive female populations (23a). HPV infection is considered a necessary
butnotasufficientriskfactorfor thedevelopmentof cervical cancer(9, 24, 29). HPV type 16 and 18 DNAs arefound in mostcancerbiopsy specimens (6, 11, 17, 30),whereasmany moretypesarepresentincervical dysplasias (11, 18).
Papillomavirus infections are a diagnostic problem
be-cause HPV cannot be cultivated in vitro and viral antigens
cannotbereliablydetected in clinicalspecimens. The
pres-ence of HPV is suspected by the presence oftypical
mor-phological changes in the epithelial cells detectable by cytology or histopathology. Specific diagnosis is, however,
only possible by nucleic acidhybridization with type-specific probes(20). SeveralDNAhybridization methodsareinuse,
including Southern blot, dot blot, filter in situ, and in situ hybridization ontissue sections (21).
We have developed a solution hybridization method,
basedon sandwich hybridization followed by affinity-based
collection ofthehybrids, for the detection of microbialDNA fromcrudeclinical specimens(14, 16, 26). One suchtest,the AffiProbe HPV test kit, is aimed at the detection ofHPV DNA from genital scrapes. The specimen is divided into
three aliquots for the detection HPV type 6/11, 16, or 18
DNA, respectively. The sensitivity of the test is 5 x 105
target molecules. The results are obtained as numerical values, andthe test standards give a clearcutofffor distin-guishing positive specimens from negativeones (T. Jalava, A. Kallio, A. W. Leinonen, and M. Ranki, Mol. Cell. Probes, in press).
*Corresponding author.
The purpose of the present study was to evaluate the AffiProbe HPVtest kitin the detection and typing of HPV DNAs fromgenital scrapes and to compare the efficacy of thetestwith that ofareference method, for whichwechose thecommercially availableViraPap and ViraTypetest kits. The latter are dot blot hybridization tests in which the samples are initially screened by the ViraPap testand the positive samples are subsequently typed by the ViraType test. TheViraTypetestallows detection of HPVtypes6/11, 16/18, and 31/33/35. Thetest results arevisualized by
auto-radiography. For interpretation of the ViraPap test, the signal intensity is compared with those of positive controls. Comparison of thetestefficacieswasrestrictedtoanalysis of cervical scrapes forwhichpaired specimens wereavailable and for the presence of HPV types 6/11, 16, and 18. The DNA tests had similar sensitivities, allowing detection of HPV DNAs from 63% ofthe cervical scrapes with benign
atypiaordyskaryosis.
MATERIALSANDMETHODS
Patients. A total of 178 patients attending the outpatient clinic of the Department of Obstetrics and Gynecology, University Central Hospital, Helsinki, Finland, were
in-cluded in thestudy. The patients belonged tothe following categories: 27 patients from the colposcopy clinic, 94
pa-tients from the emergency room, and 57 patients from the
abortion clinic.
Specimens. All patients were subjectedtoagynecological
examination during whichacervicovaginal cytologic smear
(Papanicolaou smear) was obtained. Smears were stained withamodified Papanicolaoustain.Twocervical specimens, both containing endocervical and ectocervical cells, were
collected from each patient for the DNAtests, one with a
cytobrush by using the AffiProbe specimen collection kit (OrionCorp., Orion Pharmaceutica,Helsinki, Finland) and 2076
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the other with a swab by using the ViraPap specimen collection kit (Life TechnologiesInc., Gaithersburg, Md.). The Papanicolaou smear was always taken first, but the orderof collection of the specimens for the DNA tests was randomized. The AffiProbe specimens were processed dur-ing the same day to recover the cells from the collection medium by centrifugation for 5 min at 850 x g. The cells, with100,ul of supernatant, were kept frozen until they were examined. The ViraPap specimens were also frozen until they were studied. A similar scrape from the vaginal epithe-lium of all patients was collected for the AffiProbe test. Additional specimens for the ViraPap and ViraType tests wereavailablefrom the vaginas of 65 patients and the vulvas of54 patients. The specimens were subjected to analysis with theAffiProbeHPV identification kit (Orion Corp.) and the ViraPap and ViraType tests (Life Technologies Inc.), respectively. Comparisonof the test efficacies was restricted toanalysis of174 paired cervical specimens.
AffiProbe HPV identification test. The AffiProbe HPV test kit allowsfor thesimultaneous detection and typing of HPV types 6/11, 16, and 18. For that purpose, the AffiProbe specimen was divided into three 30-pÀl aliquots, each of which was treated separately with the specimen pretreat-ment solution to release and denature the viral DNA. The aliquots were subjected to hybridization with type-specific detector probes and the biotinylated capture DNA. The probes werelabeledwith"Sisotope. After hybridization for 3 h at65°C, the hybridization mixtures were transferred to microdilution wells coated with streptavidin. Binding was for2h at37°C.The plates were then washed, and the labeled DNA that bound to hybrids was eluted with the elution solutionandsubjectedtocountingfor 5 min in a scintillation counter (LKB Wallac, Turku, Finland). Positive and nega-tivecontrolswereincludedin each type-specific test series: three controls for background, two low-positive controls, andtwohigh-positivecontrols. The cutoff of the test was set accordingto themean of the low-positive standards, which, in all cases, was at least 1.5 times the mean of the back-ground,i.e., negative control values. Eight test series were carried out.Themeanof the cutoff-to-background ratio was 2.5 (range, 1.6 to4.1) for HPV type 6/11-, 4.5 (range, 2.4 to 8.3) for HPV type 16-, and 6.0 (range, 1.9 to 11) for HPV type 18-specific tests.
ViraPap and ViraType tests. ViraPap and ViraType tests (LifeTechnologies Inc.) areintendedfordetection of DNAs ofHPV types 6/11, 16/18, and 31/33/35, as groups, by using probemixtures labeledwith32P isotope. The specimen DNA was dotted onto a nylon membrane and hybridized with a pool ofthe probemixtures described above (ViraPap test). ViraPap-positive specimens were subsequently tested as three separate dots by using three group-specific probe mixtures in thehybridizations (ViraTypetest). Resultswere visualized by
autoradiography.
Typing results of positive specimens requirefrom 4days to 2 weeks, depending on the age ofthe probe.EnzymaticamplificationofHPV DNA by usingtype-specific primers (PCR). For the polymerase chain reaction (PCR) (13), DNA was isolated from the remaining aliquot of the AffiProbe specimen by phenol extraction and ethanol pre-cipitation. The resulting DNA was dissolved in H20. PCR analysis forHPV types 6, 11, 16, and 18 usedtype-specific biotinylated primers followed by quantification in a hybrid-ization test as described previously (8a, 25). All primers weredesigned accordingtopreviously published sequences (3, 4, 22, 23). As controls, weused linearized recombinant plasmidDNAscontainingtheappropriateHPV genome and
U.
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A Positive specimens
;
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Negativespecimens
O-HPV6/11 HPV1M HPV18
'o~~~~~~~~~~~~~~~~~~~f
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LL 0.0 1. 11 UO .v .L LIO m
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RATIO (TESTRESULT/CUT OFF)
FIG. 1. ThepresenceofHPVtype6/11, 16,and 18DNAsin352
specimenswasassessedbyusing the AffiProbeHPVidentification kit. Theratio ofeachtestresulttoitsrespectivetype-specific cutoff valuewascalculated.(A)Positiveresults (ratio, 21)for HPVtypes
6/11, 16, and 18 aredepicted above the cutoSline. The means standard deviations(verticalbars)of the negativeresults (ratio,<1)
arebelow thecutoffline. Note that the scalebelowthecutoffline is linear. Thenumber of negative resultswas 328for HPVtype6/11, 324for HPVtype16,and 333 for HPVtype18. (B)Distributionsof theratios of thenegativetestresults.
pools of positive and negative patient samples from which the DNAwas purified. A sensitivity similar to that of the AffiProbe test was obtained with
104
molecules of controlplasmid DNA inthe reaction. A sample was interpretedas
positiveifthecountsperminuteobtainedafterhybridization
were the same as or higher than those obtained with the
controlplasmid.
RESULTS
PerformanceoftheAffiProbeHPVtestkit. Altogether,178
cervical and 174 vaginal scrapes were analyzed for the
a
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TABLE 1. Detection of HPV DNA from the cervical and vaginal scrapes from 178 patients by theAffiProbeHPV test Cervical No. of vaginal specimenresults
specimen result + - ND"
+ 15 20 1
- 13 126 3
a ND, Not done.
presence of HPV type 6/11, 16, and 18 DNAs by the AffiProbeHPV test kit.The cutoffvalue in each test series was defined according to the respective low-positive stan-dard. Theratio of eachtype-specifictestresult to thecutoff value was calculated. The results are shown in Fig. 1. Altogether,24,28,and 19positiveresults wereobtained for HPV types 6/11, 16, and 18, respectively. In 39% of the positive tests thesignal-to-cutoifratio was 1 to2, in44% it was between 2.1 and 10, andin17% itwasover 10(Fig. 1A). Of 985 negative tests, 867 (88%) gave counts-per-minute values beloworequaltohalf ofthecutoffvalue (Fig. 1B).
Thepositivetest results (seeabove)werederivedfrom 36 cervical and28 vaginal scrapescollected from atotal of 49 patients. In 15 (31%) patients both the cervical and the vaginalscrapeswere positive,in20patientsonly thecervical scrape was positive, and in 13 patients only the vaginal scrape waspositivefor HPV DNA (Table 1). Double infec-tions were found in seven specimens (Table 2). In one patient (patient3), thedoubleinfectionwasdetectedinboth cervical andvaginalscrapes,andinfourpatients,oneof the HPV types was also found in a second site (vagina or cervix).
Correlation between theAffiProbetestand thecontroltest in the detection of HPV DNA. The ViraPap and ViraType tests served as a reference method. For the comparison study, two simultaneously obtained cervical scrapes were available from 174 patients. A total of 18 specimens were positive and 129 specimens were negative for HPV types 6/11,16, and 18by bothtests(Table3).Thus, the agreement between the two tests was 85%. Eleven AffiProbe-positive specimens were negative by the control test. Seven addi-tional AffiProbe-positive specimens (five positive for HPV type 16 or 18 andtwo positive for HPV type 6/11; see Table 5) were suggested to contain HPV type 31/33/35 by the controltest. Eighteenadditional specimens that were
nega-TABLE 2. Double infections detected from cervical and vaginal scrapesby the AffiProbeHPV test
Test result Patient Spcmn
no. Specimen HPV HPV HPV
type 6/11 type 16 type 18
1 Cervix - + +
Vagina - - +
2 Cervix - +
-Vagina + +
-3 Cervix + +
-Vagina + +
-4 Cervix + -
-Vagina + +
5 Cervix - + +
Vagina - - +
6 Cervix
Vagina + - +
aSpecimenswithdouble infections are in boldface.
TABLE 3. Detection of HPV DNA from cervicalscrapesby AffiProbe HPVcompared with that by the controltest AffiProbe HPV No. of scrapes by the ViraType test
testresult for Toa
HPV type
6/11,
no.of HPVtype HPVtype HPVDNA 16,or18 DNA scrapes 6/11,16/18 31333 negativeDNApositive DNApositive
+ 36 18 7 il
- 138 9 18 111a
aIncludeseight scrapesthat werepositive bytheViraPaptestonly.
tive by the AffiProbe test contained HPV type 31/33/35 by the controltest(probesfor HPVtypes31, 33,and 35 are not included in the AffiProbe HPVtest).
HPV DNA in relation tocytologie findings. APapanicolaou smear wastaken fromeachpatient duringthesamevisit that scrapes for the DNA tests were obtained. HPV DNA was detected by either test in 23% of women with normal cytologyand in63% ofpatientswith abnormalcytology,i.e., Papanicolaou smear findings consistent withbenign atypia (class 2) or cervicaldysplasia (class 3) (Table 4).
Analysis of discrepant results. Independent control tests were performed on the 18 AffiProbe-positive and the 9 ViraType-positive clinical specimenswithdiscordantresults (Tables5and6). These control tests included a PCR for the detection ofHPV DNA from the remaining original speci-men, when available, detection of HPV DNA from the vagina or vulva, and repeat cervical scrapes collected on reexamination of patients who were not yet treated. The cervical scrapewas divided intotwoidentical aliquots, one fortheAffiProbeHPVtestand theother for theViraPap and ViraTypetests.
ForPCR, materialwasavailablefrom17of27specimens. Of these, 6 were
positive
and 11 werenegative
for HPV DNA (Tables5and6). In 2specimens, the amountof DNA was at least 500 ng, andin15 specimens itwasless than 250 ng. In the groupofspecimenswith alowyield of DNA, four specimens were positive, whereas both specimens with a substantialquantity ofDNA were positive.Of11AffiProbe-positive but ViraType-negative specimens (patients 1 to 11, Table 5), only 5 were available forrepeat sampling.Threeofthese werepositive. HPV DNA was also detectedineitherthevaginaorvulvasamplefromtwoof the patients.ThePapanicolaousmear wasabnormal,suggesting HPVinfection in four patients (Table 5).
Of nine ViraType-positive but AffiProbe-negative speci-mens(patients 19 to 27; Table 6) only one wasavailable for repeatsampling (patient 21), but the specimen wasnegative. HPV DNA was present in vaginal specimens from five patients,and thePapanicolaousmear suggested HPV infec-tion infour patients.
Ofsevenpatientswithdiscrepanttypes(patients 12 to 18; Table 5), patient 12 waspositive for HPV type 6/11 DNA, as
TABLE 4. Correlation of presence of HPV DNA to cytologyincervicalscrapes
Cytology Totalno. HPV DNAa
Cyof patients No. (%) positive
No.
(%)negative
Normal 122 28 (23) 94 (77)
Benignatypia 43 25 (58) 18 (42)
Dyskaryosis 13 10 (77) 3 (23)
aResults obtained by theAffiProbe and ViraType assays are combined.
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TABLE 5. Specificity ofAffiProbeHPV test
Original scrape HPVtype HPV typeon reexamination by:
Patient concurrently
no.' AffiProbe PCR Papanicolaou detected at other AffiProbe ViraType
(HPV type) (HPVtype) smearresult genital site
1 6/11 N NDd ND
2 6/liC N ND ND
3 6/11e BA 6/11, 16
4 16 16 N
5 16 16 BA 16/18
6 16 ND N ND ND
7 16C D 16/18 16
8 16C ND N 16 ND ND
9 18 ND BA ND ND
10 18 18 N ND ND
il 18C N
12 6/11 6/11 N ND ND
13 16C N ND ND
14 16C + 18C 16 BA 18 16e 16/18e
15 6/11 ND BA 31/33/35 31/33/35
16 16 BA 31/33/35 ND ND
17 16 ND D 31/33/35 ND ND
18 18 D 31/33/35 ND ND
a Patients1through 11 had a cervicalspecimenpositive by the AffiProbe HPV test butnegative by theViraType assay. Specimens from patients 12 through 18 were positive by theAffiProbeHPVtestandweretyped to containHPVDNAofHPVtypes31,33,and 35bythecontroltest.
bN,Normal; BA,benign atypia; D, dyskaryosis.
C Ahighcounts-per-minute valuewasobtained withasignal-to-cutoffratio of
.2.
dND,Not done.
eResults for thevaginal specimenonreexaminationareshown. Theconcurrentcervical scrapewasnegative bytheAffiProbe andViraTypetests.
confirmedbyPCR. Inpatient 13, the signal in the HPV type 16 DNA test was high. Patient 14 had a double infection with HPVtypes 16 and18 bytheAffiProbe test, and the presence of HPV type 16 DNA wasconfirmed byPCR. HPV type 18 was also detected in the vagina. On reexamination, HPV type 16DNA was detected by both tests. In the other four patients (patients 15 to 18), the AffiProbe test showed low counts per minute. In three patients, DNA of HPV type 31/33/35 was also found in the vulval specimen, and in the fourth patient it was found in the vaginal specimen. In one patient (patient 15), the presence of HPV type31/33/35 was confirmed by repeat sampling.
Inconclusion, independentevidence, including cytology, supported the specificity of the AffiProbe HPV test in specimensfrom 7of 11 patients (64%) that were negative by the ViraType assay. The specificity of the ViraType assay was suggested by similar analysis of specimens from seven
TABLE 6. Specificity ofthe ViraType test
Original scrape HPVtype
con-Patient currently
de-no.a ViraType PCR Papanicolaou tected atother (HPV type) (HPVtype) smearresult genitalsite
19 6/11 N 6/11
20 16/18 18 BA
21 16/18 N 16/18
22 16/18 N
23 16/18 NDC BA
24 16/18 ND N 16/18
25 16/18 ND N
26 16/18 ND BA 16/18
27 16/18 ND D 18
aThecervical scrapesfrom patients 19 through27werepositive by the ViraType assaybutnegative bytheAffiProbeHPVtest.
b N,Normal; BA,benignatypia; D,dyskaryosis.
cND,Notdone.
of nine
patients
(78%)
that werepositive
by
theViraType
testbut
negative
by
theAffiProbetest.Of
thesevenpatients
with
discrepant specimen typing results,
independent
evi-dencesupported
the AffiProberesult intwo and HPV type 31/33/35 DNA waspresentinfour.DISCUSSION
The AffiProbe HPV test kit was
developed
to achieve astandardizable, easy-to-perform
DNAhybridization
test suitable for use in anylaboratory.
Thespecimens
require
minimalpretreatment, and becausethereaction takes
place
insolution,
hybridization
is short and crudespecimens
canbe tested without
background problems.
The testyields
numerical
counts-per-minute values,
allowing
unambiguous
interpretation
ofthe results.Type-specific
internal
positive
standards define the
cutoff
values. Tostudy
the clinicalapplicability
ofthe new AffiProbe HPVtestkit,
352genital
scrapes wereexamined.
Overall,
288specimens
werenega-tiveforHPV
DNA,
57specimens
werepositive
foroneHPVtype, and 7
specimens
werepositive
fortwoHPVtypes.
Of
the
negative
specimens,
88% had asignal
that washalf
orless than
half
ofthecutoff
value. Inonly
15tests(1.5%),
thesignal
was between 0.8 and 0.99 times thecutoff
value(Fig.
1).
Erythrocytes
or mucus that werefrequently
present
inclinical
specimens
did not affectthesignal
levels.Thus,
thenegative
results wereclearly
distinguishable
from the posi-tive ones.Ithas
previously
beendocumented that unrelated bacteriaor viruses do not cause
background problems
in the Af-fiProbe HPVtest(Jalava
etal.,
inpress).
Nocross-reactions between HPV type6/11,
16,
or 18 DNA occurred in thetype-specific
tests. Aquantity
of about 109 unrelated HPVDNA molecules was
required
for apositive
signal.
In thisstudy,
thequantity
ofHPV DNA in the cervical andvaginal
scrapeswas
mostly
below107
molecules
butwasneveri109
molecules. The
specificity
of the test was studiedby
using
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crude clinicalspecimens. Specimens(11%) that gaveasignal fortwo HPV typesprobably represented truedouble
infec-tions,
and the rate was within the same order as that observed in previous studies. The HPV type 16 reagent of the AffiProbetest kitdetected 1.5 x 107 molecules of cloned HPV type 31 DNA (data not shown) because of the 70% sequencehomologybetween HPV type 16 and 31 DNAs(7). This cross-reaction apparently accounted for the four falsetyping
results obtained bythe AffiProbe test. Nevertheless,a standardized test with quantifiable numerical results is
likely
toyield
typing results equally reliable as those ob-tainedbySouthernblotting, whichis hampered by technicalas well as interobserver interpretation problems between laboratories (1).
We chose another commercially available test (the Vira-Pap and ViraTypetests) foruseas a reference method. The AffiProbetestandthedot blot test were assumed to detect a similar
quantity
ofHPV DNA, with the sensitivities of the testsbeing
unaffected by the numberof viral DNA copies percell.Agreementas tothe presenceof HPV type 6/11, 16,or 18 was obtained in 85% of the specimens, and the sensitivities of both tests for HPV types 6/11, 16, and 18 were indeed comparable. The ViraType test has probes for HPVtypes31, 33,and 35; but theseprobes are not included in the AffiProbe test. Therefore, its overall sensitivity in
detecting
HPV infections was higher than that of the Af-fiProbe test. Control tests or other independent evidenceapplied
to analysis of the discrepant results suggested that both DNAtests are specific.Previous studies have shown variable agreement as to HPV DNA positivity in concurrently collected separate
specimens
(2, 8). Furthermore, the HPV DNA detection rate iscritically affected by thenumberof cells collected (12, 15,28).
Wetherefore feel that the mostplausible explanation for thediscrepant
results in this study was an uneven distribu-tion of exfoliated cells intothe two separate specimens. Thespecimens
were not pooled and divided into two aliquots,because,
according to the instructions of the test manufac-turer, sampling is considered partof the identification kit.In this study, HPV type 6/11, 16, 18, 31, 33, or 35 DNA
was present in 63% of the cervical scrapes collected from women with cytologic atypia. The HPV DNA positivity as correlated with cytologic atypia was in the same range as that obtained by using Southern blotting for studying exfo-liatedcells (19, 28). By studying vaginalscrapes, inaddition to cervical scrapes, for the presence of HPV DNA, addi-tional information is obtained. Genital HPV infections seem
to be multifocal (10, 18). It was recently reported (28) that theaddition of cells fromtheposterior vagina to the cervical
specimen
increasedthe rate of HPV DNA detectionsignifi-cantly.
The presence of HPV DNA detected by such com-binedspecimens
couldregularly be confirmed by concurrent cervicalpunchbiopsy (19).For our study, defined groups of mostly young females
were selected. In these groups the incidence of HPV infec-tions was high and the observed high number of
false-negative
Papanicolaou smears wasthereforenotunexpected(5,
6, 27). By collecting multiple genital scrapes, the total number of HPV DNA-positive patients increased from 63 to 76 (Table 1). The rate of HPV DNA-positive patients was 33% (31 patients) in the group of women visiting the emer-gency room, 60% (34 patients) in the patients visiting the abortionclinic,and 40% (11 patients) in the patients from thecolposcopy
clinic. Unexpectedly, the number of patients withcytologicalsigns of HPV infection in the last group was small(37%).
All HPV DNA-positive women with cervicaldysplasia (10 patients) had
positive
cervical scrapes, but specimens from othergenital
sites were alsofrequently
positive (Tables 5 and 6).
The sensitivitiesof the conventional HPV DNA detection methods areapparently sufficient for
detecting
HPV-associ-ated lesions. When combined with aPapanicolaou
smear, HPV DNA detection increases theprobability
ofpredicting
histologically proven HPV lesions (19). An
important
ques-tion in thediagnosis
ofHPVinfection isthestandardization of the assays toimprove
the interobserverreproducibility
betweendifferent studies(1).
Weconcludethat theAffiProbe HPV test is a standardized hybridizationtest which is easy toperform, and theresultsare availablein 1working
day.
It has asensitivitycomparable to orslightly higherthan thatof the ViraType test indetecting
HPV type6/11, 16,
and 18 DNAs.ACKNOWLEDGMENTS
We thank Seija Lehto for excellent technical assistance,
Marja-LiisaOgàrd for helpincollecting the specimens,andHeiniJarvifor processing themanuscript.
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