Research Article The Prevalence of Human Papillomavirus

Research ArticleThe Prevalence of Human Papillomavirus betweenthe Neonates and Their Mothers

Mariusz SkoczyNski1 Anna Gofdzicka-Joacutezefiak2 and Anna KwaVniewska1

1Department of Obstetrics and Pathology of Pregnancy Medical University of Lublin 20059 Lublin Poland2Department of Molecular Virology Institute of Experimental Biology Adam Mickiewicz University 61712 Poznan Poland

Correspondence should be addressed to Mariusz Skoczynski mskoczynskiwppl

Received 20 August 2015 Revised 3 November 2015 Accepted 19 November 2015

Academic Editor Adriana M Vieira

Copyright copy 2015 Mariusz Skoczynski et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

The impact of human papillomavirus (HPV) infection on pregnancy is a major problem of medicineThe transmission of the virusfrom mother to fetus is a process yet unresolved The immune response and changed hormonal status of pregnant women mightfacilitate infection A research on the prevalence of HPV infection was conducted at the Clinic of Obstetrics Medical Universityof Lublin (Poland) The studied group included 152 randomly selected women The material was tested for the presence of HPVDNA by means of polymerase chain reaction (PCR) The aim of the research was to assess the relation between HPV infectionsdetected in the buccal smears of the neonates and the incidence of such infections in the cervicalbuccal smears of their mothersIn the group of 152 infants HPV was found in 16 (1053) Among the cervicalbuccal smears HPV was isolated respectively in 24(1579) and in 19 (125) pregnant women Statistically significant differences in the prevalence of HPV swabs from the newbornsand the cervicalbuccal smears of their mothers were found (p lt 0001)The identification of mothers in whose buccal smears HPVwas detected can help develop a group of children who run a relatively significant risk of being infected

1 Introduction

The influence of human papillomavirus infection on preg-nancy remains an unresolved matter HPV infection is com-monly included in the group of sexually transmitted diseases(STDs) and is one of the most frequently transmitted amongthem [1 2] The influence of the high-risk HPV virus on can-cerous processes is a commonly accepted hypothesis [3] Therelation between HPV infection and the development of RRP(Recurrent Respiratory Papillomatosis) in newborns cannotbe omitted According to the various reports 30 to 50 ofthe clinical signs of the infection in mucous membrane ofthe upper respiratory tract caused by human papillomaviruscan be associated with motherrsquos HPV infection [4ndash6] Theestimated risk ratio of infection among young women is80 although the majority of infections are asymptomaticand show a tendency to cure spontaneously [7 8] It shouldhowever be underlined that women aged between 20 and 35not only are most prone to getting infected with HPV butalso are most sexually active get pregnant and have and raise

children The possibility of HPV transmission from motherto fetus occurring during pregnancy and perinatal periodhas been postulated It is however still undetermined andsparks heated debate Therefore identifying the risk factorsand mechanisms of transmission of the virus includingthe impact of hormonal and immunological changes duringpregnancy is of vital importance Taking into considerationthe current epidemiological data it can be concluded thatHPV transmission from mother to newborn is possiblebefore during and after birth [9ndash11] It confirms the impor-tance of the transmission of the virus from mother to herbaby The presented study aims to compare the frequencyof HPV infection in pregnant women and their newbornsin different places An attempt to find correlation betweenmothersrsquo and newbornsrsquo infectionswasmade to select a groupof children prone to HPV-related entities Apart from thecytological examination of cervix the smear of oral mucosaboth frommothers and their babies used for the identificationand prophylaxis ofHPV infection could be relevant in clinicalpractice

Hindawi Publishing CorporationBioMed Research InternationalVolume 2015 Article ID 126417 6 pageshttpdxdoiorg1011552015126417

2 BioMed Research International

2 Materials and Methods

This study was conducted between March 2009 and June2011 and included 152 pregnant women who delivered at theClinic of Obstetrics and Pathology of Pregnancy MedicalUniversity of Lublin (Poland) The studied group includedrandom obstetric population recruited prior to the deliveryThe routine medical examination was followed by previoushistory of HPV-related genital diseases and treatments Thestudy had the following inclusion criteria (1) singleton preg-nancy (2) lack of clinical symptoms suggesting the possibilityof HPV infection (3) normal cervical smear and (4) termdelivery The number of women who refused was 17 Theexclusion criteria included (1) history of HPV infection (119899 =7) (2) abnormal cervical smear (119899 = 4) and (3) multiplepregnancy (119899 = 6) The subjects gave their written informedconsent before the start of any procedure The protocol ofthis study was approved by the Local Bioethical Committeeat the Medical University of Lublin Prior to delivery smearswere obtained from the cervix and buccal mucosa of eachpregnant woman The cervical HPV DNA specimen wascollected with a cytobrush inserted into the vagina Thecervix and ectocervix were swabbed using a circular motionThe oral specimens were collected during the same timeImmediately after the birth the neonatal oral mucosa smearswere obtained in similar proceedings with a cytobrush Allsamples were collected by the same properly trained personThematerial was placed in sterile tubes for HPVDNA testing(Eurotubo Deltalab Spain) frozen and stored at 70∘C untilfurther analyses The material was tested for the presenceof HPV DNA by means of PCR method Identification ofHPV types was performed using consensus PCR primersfor L1MY09 51015840-CGTCCMARRGGAWACTGATC-31015840 andMY11 51015840-GCMCAAGGWCATAAYAATGG-31015840 where M =A+C R=A+GW=A+T andY=C+TThis set of primersamplifies DNA from at least 33 different HPV genotypesHPV types 16 andor 18 were identified using the fol-lowing type specific PCR primers HPV16L1AHPV16L1B51015840-GCCTGTGTAGGTGTTGAGGT-31015840 and 51015840-TGGATT-TACTCCAACATTGG-31015840 product size 264 bp HPV18L1AHPV18L1B 51015840-GTGGACCAGCAAATACAGGA-31015840and 51015840-TGCAACGACCACGTGTTGGA-31015840 product site162 bp HPV18ME12HPV18ME50E6 51015840-CACGGCGAC-CCTACAAGCTACCTG-31015840 and 51015840-TGCAGCACGAAT-TGGCACTGGCCTC-31015840 product site 404 bp The reactionwas performed according to Tucker et al [12] The totalvolume of 10 120583L of PCR mixture contained 1 120583M of primers200120583M of deoxynucleotide triphosphates 1x PCR buffer(01M Tris-HCl pH 88 05M KCl 0015MMgCl

2 1 Triton

X-100) the investigated DNA (10 ng120583L) and Tag poly-merase at a final concentration of 40UmL After preliminarydenaturation (15min at 94∘C) samples were amplified for31 cycles in a thermal cycler Each cycle consisted of thefollowing steps denaturation at 94∘C for 30 seconds andannealing at 59∘C for the same amount of time followed byprimer extension at 72∘C for 1min In the last PCR cyclethe stage of complementary DNA synthesis at 72∘C wasextended to 420 seconds PCR products were analysed usingagarose gel electrophoresis in the presence of pBluescript

DNA digested with HindI The presence of HPV DNA wasadditionally verified and HPV genotypes were identified bydirect sequencing from CR reaction tube (without purifi-cation from MY09 and MY11 oligonucleotides) The resultsof sequencing were analysed using the BLAST database(httpblastncbinlmnihgov)

21 Statistical Analysis Normal distribution of continuousvariables was verified with the Kolmogorov-Smirnoff testStatistical characteristics of these variables were presented asarithmetic means and standard deviations (SD) and theirintergroup comparisons were performed with Studentrsquos 119905-testfor independent variables All calculations were carried outwith Statistica 10 (Stat Soft Tulsa OK USA) package withthe level of significance set at 119901 le 005

3 Results and Discussion

31 Incidence of HPV Infection in the Oral Mucosa of Motherand Her Child As the result of the conducted research HPVDNA was isolated in 125 (19152) of swabs from the oralmucosa of the mothers and in 1053 (16152) of the childrenIn the group of nineteen mothers with HPV infection wefound 8 (4211) HPV-positive children and 11 (5789)HPV-negative children In the mothers group without HPVinfection 133152 (875) we observed 8133 (602) HPV(+) newborns and 125133 (9398) HPV (minus) newborns Astatistically significant difference in the prevalence of HPVDNA in the swabs from the oral mucosa of the newbornsand the swabs from the oral mucosa of their mothers wasfound (plt 0001)There is an eleven times greater relative risk(OR = 11363 CI 3536ndash36509) of discovering HPV DNA inbuccal smears of newborn of a HPV-positive mother than ofthe mother with negative results of HPV The correlation ofthe frequency of HPV infections was presented in Table 1

The idea of HPV transmission from mother to child wasfirst introduced by Sedlacek et al [13] in 1989 Dependingon the population tested the place where the tested materialis obtained and methods used to identify HPV DNA thefrequency of HPV transmission varies from 4 to 80 [1415] According to Bandyopadhyay et al such a wide range ofgathered results is also a proof of temporary contaminationwith the infected material [16] Cason et al suggested thatbuccal smears provide better material for analysis of HPVDNA incidence than respiratory discharge samples [17]However in contrast to our data Eppel et al presentedapproximately two times higher results in their PCR studyThe prevalence of HPV DNA in specimens from cervicalsmears of asymptomatic pregnant women was found in 246 (44179) [18] Such a confusing difference may be dueto several confounders including study design choice ofdetection methods and differences in risk factors for HPVinfection Another researchwas conducted in 2005 in Finlandwhen the first prospective tests of HPV infection incidencewere ordered among family members This study analysedthe relationship between the presences of HPV infection infamily environment The team of researchers led by Rintalafocused on a group of 76 families and recognized HPVinfection in 8 of buccal smears of mothers and 10 of

BioMed Research International 3

Table 1 The correlation of the frequency of HPV infections between buccal smears (119899 = 152)

Newborn buccal smears Total number ()1HPV DNA (+) HPV DNA (minus)

Mother buccal smears HPV DNA (+) 8lowast 11 19 (125)Mother buccal smears HPV DNA (minus) 8 125 133 (875)Total number ()2 16 (1053) 136 (8947) 152 (100)1Mother buccal smears2Newborn buccal smearslowast

119901 lt 0001

Table 2 The correlation between HPV DNA incidence in oral mucosa of newborn and the cervix of mother (119899 = 152)


Mother cervical smears HPV DNA (+) 11lowast 13 24 (1579)Mother cervical smears HPV DNA (minus) 5 123 128 (8421)Total number ()2 16 (1053) 136 (8947) 152 (100)1Mother cervical smears2Newborn buccal smearslowast

119901 lt 0001

buccal smears of their neonates The results of our researchconducted on a study group twice the size show similarities[19]

32 Incidence of HPV Infection in the Oral Mucosa of Childand the Cervical Smears of Mother In the studied group(119899 = 152) HPV DNA was isolated in 24 (1579) swabsfrom the mothersrsquo cervix and in 16 (1052) from oralmucosa of newborns Out of 24 HPV-positive mothersHPV DNA was isolated in 11 (4583) of their children Ingroup of 24 HPV-positive mothers we found 13 (5417)HPV-negative children In the mothers group without HPVinfection 128152 (8421) we observed 5128 (391) HPV(+) newborns and 123128 (9609) HPV (minus) newborns Astatistically significant difference in the prevalence of HPVDNA in the swabs from the oral mucosa of the newbornsand the swabs from the cervix of their mothers was found(p lt 0001) Based on the analysis of the results the relativerisk of HPV DNA existing in swabs from the oral mucosaof the children whose mothers were tested positive for HPVin cervical smears is almost 21 times greater than in childrenwhose mothers were diagnosed as healthy (OR = 20815 CI6197ndash69914)The correlation between HPV infection in oralmucosa of children and the cervix of mother was presentedin Table 2

Similar results were obtained by Rice et al [20] as HPVDNA was found in 30 of the children whose motherswere infected They concluded that mothers with high HPV-16 viral loads in their genital tracts are usually the sourceof newborns infections How many newborns are infectedremains a vital question It was suggested by the authorsthat there is a correlation between the type of virus andits transmissive abilities They recognized that not enoughattention was paid to show that at least 20 of healthychildren present HPV infection connected with perinatal

time The same observation was supported by Puranenet al [15] They concluded that perinatal acquired HPVinfectionmight persist in the oral cavity for years without anysignificant clinical lesions and infected mother can transmithuman papillomavirus to her child In this study HPV DNAwas found in 31 of the 98 (316) mothersrsquo oral scrapingsOur study revealed 4583 neonatal prevalence of HPVin children whose mothers were tested HPV positive incervical smears at delivery Very similar data was presentedby Sedlacek et al [13] They demonstrated HPV DNA inthe oral cavity of 478 (1123) of the neonates deliveredvaginally from mothers in whose cervical cells HPV DNAwas detected These results are also consistent with thosereported in Puranen et al study [15] It has been suggestedthat neonates could acquire HPV infection as they passthrough an infected birth canal [13 15 20]

33 HPV Infection Incidence in Women Giving Birth HPVDNA was identified in 29 women giving birth which was1908 of the total population (119899 = 152) Among the swabsof the cervical smears HPV DNA was isolated in 24152(1579) women and among the swabs from the oral mucosaHPVDNAwas observed in 19152 (125) Both variables areseparable and analysed as such Simultaneous HPV infectionin both swabs from the oral mucosa and the cervix wasfound in 14152 (921) of the total population NoHPVDNAwas found in any of the assessed swabs taken from 123152(8092) women giving birth As a result of the research astatistically significant difference in HPV DNA incidence incervical smears in relation toHPVDNAoccurrence in buccalsmears of women was found (p lt 0001) Out of 19 women inwhose swabs from the oral mucosa HPV DNAwas identifiedthe virus was also found in 14 swabs from cervix There isa 34 times greater relative risk (OR = 34446 CI 10194ndash116353) of HPVDNA incidence in swabs from cervix if HPV


Table 3 The correlation between HPV DNA incidence in the cervicalbuccal smears of mothers (119899 = 152)

Mother cervical smears Total number ()1HPV DNA (+) HPV DNA (minus)

Mother buccal smears HPV DNA (+) 14lowast 5 19 (125)Mother buccal smears HPV DNA (minus) 10 123 133 (875)Total number ()2 24 (1557) 128 (8443) 152 (100)1Mother buccal smears2Mother cervical smearslowast

119901 lt 0001

DNA is found in oral mucosa smears of the patient Thecorrelation between HPV infections in the cervicalbuccalsmears of woman giving birth was presented in Table 3

A well-documented relation between the frequency ofHPV infections young age contraceptives intake smokingnumber of sexual partners marital status and level ofeducation obtained exists [15 21 22] A research team led byTakakuwa analysed a study group of 1183 pregnant patientsamong whom HPV infection was detected in 125 of cases[23] It is worth noticing that young patients are statisticallymore prone to HPV infections as HPV DNA was detectedin 226 of the patients under the age of 25 and 113 ofolder patients (p lt 00005) Place of residence of the patientsmight be another factor that determines the frequency ofHPV infections In pregnant and nonpregnant women inUganda HPV DNA is detected in not less than 60 of caseswhereas it is estimated that 65 of pregnant women in Spainare infected [24 25] A research conducted in a group of 291pregnant women in South Korea determined the frequencyof HPV infection to be 189 Additional stress was puton more frequent HPV infections in women with abnormalcytology with no record of pregnancies and under the ageof 30 at the time of the tests [26] Furthermore a divisionbetween pregnantwomenwithout symptoms of infection andpregnant patients with visible changes to the skin andmucosawas made In a group of pregnant patients suffering from anasymptomaticHPV infectionWatts assessed the transmissionof virus to newborns to be 1 [27] Similar results canbe found in research conducted by Kashima and coauthorswho focused on a group of 231 newborns in whose swabsfrom mucosa and genital region HPV DNA was observed[28] What is more Smith and coauthors in their researchon the incidence of HPV infection in children of infectedmothers concluded that HPV DNA was found in 18 ofswabs taken from 6-week-old children [29] Presented dataprove the existence of vertical HPV transmission althoughthe risk is considered low [18 23 30 31] However the virustransmission incidence in cases of mothers who developedsymptoms of infection is estimated to be 5 to 72 [3233] Higher relative risk of HPV infection in child whosemother is HPV (+) seems to be logical and valid [16 18 27]The dissimilarities between results obtained during medicalresearch might be dependent on the number of copies ofthe virus in the assessed study group and the employmentof a variety of methods for HPV DNA detection [33ndash35] One of the many factors of such correlation betweenthe occurrences of HPV infection in the two analysed

environments might be local defensive mechanisms Accord-ing to Smith et al infections in swabs from cervical smearswere 10 times more frequent than in swabs from buccalsmears [14] Similar trends have been noted in publicationsconcerning pregnant women [36 37] Smaller number ofdetected HPV infections in buccal smears might be causedby different defensive mechanisms active in that particularenvironment The microbiological protection of oral cavityand more so the saliva might be crucial to the defenceagainst infection risk factors [38ndash40] There were other studylimitations The studied group could be more numerous dueto little prevalence of infected participants It is difficult toreliably verify the history of infections because some mightbe unidentified However the results of the research workcould be relevant in clinical practice The analysis of theincidence of infection of human papillomavirus HPV in fam-ily environment showed an important and often overlookedsignificance of other test methods used for the identificationof HPV infection It seems that the introduction of a readilyavailable and comfortable oral mucosa swab test should bemuch more promoted and encouraged The identification ofmothers in whose swabs of the oral mucosa HPV DNA wasdetected can help develop a group of children who run arelatively significant risk of being infected Further studiesmay be needed to monitor the development of newbornsinfections

4 Conclusions

A connection between buccal HPV incidence in mothersand newborns was proved in the research A statisticallysignificant difference in HPV DNA incidence in cervicalsmears in relation to HPV DNA occurrence in mothersrsquobuccal smears was found A thorough examination andfollow-up of HPV (+)mothersrsquo newbornsmay be vital for theselection of a group of children prone toHPV-related entities

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This work was supported by Found Grand KBN no NN407016135 provided by the Polish Ministry of Science and


Higher Education The authors acknowledge Dr MarcinSkoczynski for his assistance in editing the paper

References

[1] M Merckx W Van Wouwe Liesbeth M Arbyn et al ldquoTrans-mission of carcinogenic human papillomavirus types frommother to child ameta-analysis of published studiesrdquo EuropeanJournal of Cancer Prevention vol 22 no 3 pp 277ndash285 2013

[2] H Zur Hausen ldquoPapillomavirus infectionsmdasha major cause ofhuman cancersrdquo Biochimica et Biophysica ActamdashReviews onCancer vol 1288 no 2 pp F55ndashF78 1996

[3] IARC Human Papillomaviruses vol 90 of IARC MonographsInternational Agency for Research on Cancer (IARC) LyonFrance 2007

[4] C S Derkay and B Wiatrak ldquoRecurrent respiratory papillo-matosis a reviewrdquo Laryngoscope vol 118 no 7 pp 1236ndash12472008

[5] T Q Gallagher and C S Derkay ldquoPharmacotherapy of recur-rent respiratory papillomatosis an expert opinionrdquo ExpertOpinion on Pharmacotherapy vol 10 no 4 pp 645ndash655 2009

[6] V Gerein S Schmandt N Babkina N Barysik W Coerdt andH Pfister ldquoHuman papilloma virus (HPV)-associated gyne-cological alteration in mothers of children with recurrent res-piratory papillomatosis during long-term observationrdquo CancerDetection and Prevention vol 31 no 4 pp 276ndash281 2007

[7] M H Einstein J T Schiller R P Viscidi et al ldquoClinicianrsquosguide to human papillomavirus immunology knowns andunknownsrdquoTheLancet Infectious Diseases vol 9 no 6 pp 347ndash356 2009

[8] I H Frazer ldquoInteraction of human papillomaviruses with thehost immune system a well evolved relationshiprdquo Virology vol384 no 2 pp 410ndash414 2009

[9] X Wang J Zhuang K Wu R Xu M Li and Y Lu ldquoHumansemen the biological basis of sexual behaviour to promotehuman papillomavirus infection and cervical cancerrdquo MedicalHypotheses vol 74 no 6 pp 1015ndash1016 2010

[10] Q-J Wu M Guo Z-M Lu T Li H-Z Qiao and Y KeldquoDetection of humanpapillomavirus-16 in ovarianmalignancyrdquoBritish Journal of Cancer vol 89 no 4 pp 672ndash675 2003

[11] M E Sarkola S E Grenman M A M Rintala K J Syrjanenand S M Syrjanen ldquoHuman papillomavirus in the placentaand umbilical cord bloodrdquo Acta Obstetricia et GynecologicaScandinavica vol 87 no 11 pp 1181ndash1188 2008

[12] R A Tucker P R Johnson W C Reeves and J P IcenogleldquoUsing the polymerase chain reaction to genotype humanpapillomavirus DNAs in samples containing multiple HPVsmay produce inaccurate resultsrdquo Journal of Virological Methodsvol 43 no 3 pp 321ndash333 1993

[13] T V Sedlacek S Lindheim C Eder et al ldquoMechanism forhuman papillomavirus transmission at birthrdquoAmerican Journalof Obstetrics and Gynecology vol 161 no 1 pp 55ndash59 1989

[14] E M Smith J M Ritchie J Yankowitz et al ldquoHumanpapillomavirus prevalence and types in newborns and parentsconcordance and modes of transmissionrdquo Sexually TransmittedDiseases vol 31 no 1 pp 57ndash62 2004

[15] M Puranen M Yliskoski S Saarikoski K Syrjanen and SSyrjanen ldquoVertical transmission of human papillomavirus frominfectedmothers to their newborn babies and persistence of thevirus in childhoodrdquoAmerican Journal of Obstetrics andGynecol-ogy vol 174 no 2 pp 694ndash699 1996

[16] S Bandyopadhyay S Sen L Majumdar and R ChatterjeeldquoHuman papillomavirus infection among indian mothers andtheir infantsrdquo Asian Pacific Journal of Cancer Prevention vol 4no 3 pp 179ndash184 2003

[17] J Cason J N Kaye R J Jewers et al ldquoPerinatal infection andpersistence of human papillomavirus types 16 and 18 in infantsrdquoJournal of Medical Virology vol 47 no 3 pp 209ndash218 1995

[18] W Eppel C Worda P Frigo M Ulm E Kucera and K Czer-wenka ldquoHuman papillomavirus in the cervix and placentardquoObstetrics and Gynecology vol 96 no 3 pp 337ndash341 2000

[19] M A M Rintala S E Grenman M H Puranen et al ldquoTrans-mission of high-risk human papillomavirus (HPV) betweenparents and infant a prospective study of HPV in families inFinlandrdquo Journal of Clinical Microbiology vol 43 no 1 pp 376ndash381 2005

[20] P S Rice J Cason J M Best and J E Banatvala ldquoHigh riskgenital papillomavirus infections are spread verticallyrdquo Reviewsin Medical Virology vol 9 no 1 pp 15ndash21 1999

[21] OMalhomme N Dutheil M Rabreau E Armbruster-MoraesJ R Schlehofer and T Dupressoir ldquoHuman genital tissuescontainingDNAof adeno-associated virus lackDNA sequencesof the helper viruses adenovirus herpes simplex virus orcytomegalovirus but frequently contain human papillomavirusDNArdquo Journal of General Virology vol 78 no 8 pp 1957ndash19621997

[22] E M Smith M A Parker L M Rubenstein T H Haugen EHamsikova and L P Turek ldquoEvidence for vertical transmissionof HPV from mothers to infantsrdquo Infectious Diseases in Obstet-rics and Gynecology vol 2010 Article ID 326369 7 pages 2010

[23] K Takakuwa T Mitsui M Iwashita et al ldquoStudies on theprevalence of human papillomavirus in pregnant women inJapanrdquo Journal of Perinatal Medicine vol 34 no 1 pp 77ndash792006

[24] X Castellsague T Drudis M P Canadas et al ldquoHuman Papil-lomavirus (HPV) infection in pregnant women and mother-to-child transmission of genital HPV genotypes a prospectivestudy in Spainrdquo BMC Infectious Diseases vol 9 article 74 2009

[25] C Banura S Franceschi L-J van Doorn et al ldquoPrevalenceincidence and clearance of human papillomavirus infectionamong young primiparous pregnant women in KampalaUgandardquo International Journal of Cancer vol 123 no 9 pp2180ndash2187 2008

[26] H Park S W Lee I H Lee et al ldquoRate of vertical transmissionof human papillomavirus from mothers to infants relationshipbetween infection rate and mode of deliveryrdquo Virology Journalvol 9 article 80 2012

[27] D H Watts L A Koutsky K K Holmes et al ldquoLow risk ofperinatal transmission of human papillomavirus results froma prospective cohort studyrdquo American Journal of Obstetrics andGynecology vol 178 no 2 pp 365ndash373 1998

[28] H K Kashima F Shah A Lyles et al ldquoA comparison of riskfactors in juvenile-onset and adult-onset recurrent respiratorypapillomatosisrdquo Laryngoscope vol 102 no 1 pp 9ndash13 1992

[29] E M Smith S R Johnson T Cripe et al ldquoPerinatal transmis-sion and maternal risks of human papillomavirus infectionrdquoCancer Detection and Prevention vol 19 no 2 pp 196ndash2051995

[30] F Pakarian J Kaye J Cason et al ldquoCancer associatedhuman papillomaviruses perinatal transmission and persis-tencerdquoBritish Journal of Obstetrics andGynaecology vol 101 no6 pp 514ndash517 1994


[31] R L Winer and L A Koutsky ldquoDelivering reassurance toparents perinatal human papillomavirus transmission is rarerdquoSexually Transmitted Diseases vol 31 no 1 pp 63ndash64 2004

[32] B D Fredericks A Balkin HWDaniel J Schonrock BWardand I H Frazer ldquoTransmission of human papillomavirusesfrom mother to childrdquo Australian and New Zealand Journal ofObstetrics and Gynaecology vol 33 no 1 pp 30ndash32 1993

[33] R L Rombaldi E P Serafini J Mandelli E Zimmermannand K P Losquiavo ldquoPerinatal transmission of human papilo-mavirus DNArdquo Virology Journal vol 6 article 83 2009

[34] J N Kaye J Cason F B Palrarian et al ldquoViral load as adeterminant for transmission of human papillomavirus type 16from mother to childrdquo Journal of Medical Virology vol 44 no4 pp 415ndash421 1994

[35] S Alberico R Pinzano M Comar et al ldquoMaternal-fetal trans-mission of human papillomavirusrdquo Minerva Ginecologica vol48 no 5 pp 199ndash204 1996

[36] E A B Morrison M D Gammon G L Goldberg S H Ver-mund and R D Burk ldquoPregnancy and cervical infection withhuman papillomavirusesrdquo International Journal of Gynecologyand Obstetrics vol 54 no 2 pp 125ndash130 1996

[37] M V Jacobs J M M Walboomers P J F Snijders et al ldquoDis-tribution of 37 mucosotropic HPV types in women with cyto-logically normal cervical smears the age-related patterns forhigh-risk and low-risk typesrdquo International Journal of Cancervol 87 no 2 pp 221ndash227 2000

[38] D C Shugars C A Watkins and H J Cowen ldquoSalivaryconcentration of secretory leukocyte protease inhibitor anantimicrobial protein is decreased with advanced agerdquo Geron-tology vol 47 no 5 pp 246ndash253 2001

[39] I D Miletic S S Schiffman V D Miletic and E A Sattely-Miller ldquoSalivary IgA secretion rate in young and elderly per-sonsrdquo Physiology amp Behavior vol 60 no 1 pp 243ndash248 1996

[40] C Steele and P L Fidel Jr ldquoCytokine and chemokine produc-tion by human oral and vaginal epithelial cells in response toCandida albicansrdquo Infection and Immunity vol 70 no 2 pp577ndash583 2002

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


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BioMed Research International

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Oxidative Medicine and Cellular Longevity


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The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


2 Materials and Methods

This study was conducted between March 2009 and June2011 and included 152 pregnant women who delivered at theClinic of Obstetrics and Pathology of Pregnancy MedicalUniversity of Lublin (Poland) The studied group includedrandom obstetric population recruited prior to the deliveryThe routine medical examination was followed by previoushistory of HPV-related genital diseases and treatments Thestudy had the following inclusion criteria (1) singleton preg-nancy (2) lack of clinical symptoms suggesting the possibilityof HPV infection (3) normal cervical smear and (4) termdelivery The number of women who refused was 17 Theexclusion criteria included (1) history of HPV infection (119899 =7) (2) abnormal cervical smear (119899 = 4) and (3) multiplepregnancy (119899 = 6) The subjects gave their written informedconsent before the start of any procedure The protocol ofthis study was approved by the Local Bioethical Committeeat the Medical University of Lublin Prior to delivery smearswere obtained from the cervix and buccal mucosa of eachpregnant woman The cervical HPV DNA specimen wascollected with a cytobrush inserted into the vagina Thecervix and ectocervix were swabbed using a circular motionThe oral specimens were collected during the same timeImmediately after the birth the neonatal oral mucosa smearswere obtained in similar proceedings with a cytobrush Allsamples were collected by the same properly trained personThematerial was placed in sterile tubes for HPVDNA testing(Eurotubo Deltalab Spain) frozen and stored at 70∘C untilfurther analyses The material was tested for the presenceof HPV DNA by means of PCR method Identification ofHPV types was performed using consensus PCR primersfor L1MY09 51015840-CGTCCMARRGGAWACTGATC-31015840 andMY11 51015840-GCMCAAGGWCATAAYAATGG-31015840 where M =A+C R=A+GW=A+T andY=C+TThis set of primersamplifies DNA from at least 33 different HPV genotypesHPV types 16 andor 18 were identified using the fol-lowing type specific PCR primers HPV16L1AHPV16L1B51015840-GCCTGTGTAGGTGTTGAGGT-31015840 and 51015840-TGGATT-TACTCCAACATTGG-31015840 product size 264 bp HPV18L1AHPV18L1B 51015840-GTGGACCAGCAAATACAGGA-31015840and 51015840-TGCAACGACCACGTGTTGGA-31015840 product site162 bp HPV18ME12HPV18ME50E6 51015840-CACGGCGAC-CCTACAAGCTACCTG-31015840 and 51015840-TGCAGCACGAAT-TGGCACTGGCCTC-31015840 product site 404 bp The reactionwas performed according to Tucker et al [12] The totalvolume of 10 120583L of PCR mixture contained 1 120583M of primers200120583M of deoxynucleotide triphosphates 1x PCR buffer(01M Tris-HCl pH 88 05M KCl 0015MMgCl

2 1 Triton

X-100) the investigated DNA (10 ng120583L) and Tag poly-merase at a final concentration of 40UmL After preliminarydenaturation (15min at 94∘C) samples were amplified for31 cycles in a thermal cycler Each cycle consisted of thefollowing steps denaturation at 94∘C for 30 seconds andannealing at 59∘C for the same amount of time followed byprimer extension at 72∘C for 1min In the last PCR cyclethe stage of complementary DNA synthesis at 72∘C wasextended to 420 seconds PCR products were analysed usingagarose gel electrophoresis in the presence of pBluescript

DNA digested with HindI The presence of HPV DNA wasadditionally verified and HPV genotypes were identified bydirect sequencing from CR reaction tube (without purifi-cation from MY09 and MY11 oligonucleotides) The resultsof sequencing were analysed using the BLAST database(httpblastncbinlmnihgov)

21 Statistical Analysis Normal distribution of continuousvariables was verified with the Kolmogorov-Smirnoff testStatistical characteristics of these variables were presented asarithmetic means and standard deviations (SD) and theirintergroup comparisons were performed with Studentrsquos 119905-testfor independent variables All calculations were carried outwith Statistica 10 (Stat Soft Tulsa OK USA) package withthe level of significance set at 119901 le 005

3 Results and Discussion

31 Incidence of HPV Infection in the Oral Mucosa of Motherand Her Child As the result of the conducted research HPVDNA was isolated in 125 (19152) of swabs from the oralmucosa of the mothers and in 1053 (16152) of the childrenIn the group of nineteen mothers with HPV infection wefound 8 (4211) HPV-positive children and 11 (5789)HPV-negative children In the mothers group without HPVinfection 133152 (875) we observed 8133 (602) HPV(+) newborns and 125133 (9398) HPV (minus) newborns Astatistically significant difference in the prevalence of HPVDNA in the swabs from the oral mucosa of the newbornsand the swabs from the oral mucosa of their mothers wasfound (plt 0001)There is an eleven times greater relative risk(OR = 11363 CI 3536ndash36509) of discovering HPV DNA inbuccal smears of newborn of a HPV-positive mother than ofthe mother with negative results of HPV The correlation ofthe frequency of HPV infections was presented in Table 1

The idea of HPV transmission from mother to child wasfirst introduced by Sedlacek et al [13] in 1989 Dependingon the population tested the place where the tested materialis obtained and methods used to identify HPV DNA thefrequency of HPV transmission varies from 4 to 80 [1415] According to Bandyopadhyay et al such a wide range ofgathered results is also a proof of temporary contaminationwith the infected material [16] Cason et al suggested thatbuccal smears provide better material for analysis of HPVDNA incidence than respiratory discharge samples [17]However in contrast to our data Eppel et al presentedapproximately two times higher results in their PCR studyThe prevalence of HPV DNA in specimens from cervicalsmears of asymptomatic pregnant women was found in 246 (44179) [18] Such a confusing difference may be dueto several confounders including study design choice ofdetection methods and differences in risk factors for HPVinfection Another researchwas conducted in 2005 in Finlandwhen the first prospective tests of HPV infection incidencewere ordered among family members This study analysedthe relationship between the presences of HPV infection infamily environment The team of researchers led by Rintalafocused on a group of 76 families and recognized HPVinfection in 8 of buccal smears of mothers and 10 of





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