Vaginal Microbiota Composition Correlates Between Pap Smear Microscopy and Next Generation Sequencing and Associates to Socioeconomic Status

Recent research on vaginal microbiota relies on high throughput sequencing while microscopic methods have a long history in clinical use. We investigated the correspondence between microscopic findings of Pap smears and the vaginal microbiota composition determined by next generation sequencing among 50 asymptomatic women. Both methods produced coherent results regarding the distinction between Lactobacillus-dominant versus mixed microbiota, reassuring gynaecologists for the use of Pap smear or wet mount microscopy for rapid evaluation of vaginal bacteria as part of diagnosis. Cytologic findings identified women with bacterial vaginosis and revealed that cytolysis of vaginal epithelial cells is associated to Lactobacillus crispatus-dominated microbiota. Education and socio-economic status were associated to the vaginal microbiota variation. Our results highlight the importance of including socio-economic status as a co-factor in future vaginal microbiota studies.Peer reviewe

Sisäsynnytintulehdus

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Predictive Values of Serum Chlamydia trachomatis TroA and HtrA IgG Antibodies as Markers of Persistent Infection in the Detection of Pelvic Adhesions and Tubal Occlusion

Chlamydia trachomatis IgG antibody testing (CAT) has been used as a screening test for tubal factor infertility (TFI), but as the CAT is only a marker of a past exposure to C. trachomatis and not of late sequelae, the positive predictive value (PPV) of the test is low. The persistence of C. trachomatis in the upper genital tract has been suggested as one of the key mechanisms in the development of TFI. Serum antibodies against C. trachomatis TroA and HtrA, proteins expressed specifically during persistent infection, have been suggested as novel biomarkers for TFI diagnostics. We studied serum IgG antibody responses against C. trachomatis TroA, HtrA and MOMP in 79 subfertile women, of whom 28 had laparoscopically proven TFI. We confirmed that the accuracy of CAT in diagnosing TFI is low, whereas TroA IgG and HtrA IgG are more accurate tests in detecting tubal occlusion and pelvic adhesions. However, the sensitivity and negative predictive value (NPV) of TroA IgG and HtrA IgG are still too low to justify their use as a screening test in clinical practice. Individual immunogenetic profiles combined with TroA and HtrA antibody responses might identify women with the highest risk for developing late complications after C. trachomatis infection

Predictive Values of Serum Chlamydia trachomatis TroA and HtrA IgG Antibodies as Markers of Persistent Infection in the Detection of Pelvic Adhesions and Tubal Occlusion

Chlamydia trachomatis IgG antibody testing (CAT) has been used as a screening test for tubal factor infertility (TFI), but as the CAT is only a marker of a past exposure to C. trachomatis and not of late sequelae, the positive predictive value (PPV) of the test is low. The persistence of C. trachomatis in the upper genital tract has been suggested as one of the key mechanisms in the development of TFI. Serum antibodies against C. trachomatis TroA and HtrA, proteins expressed specifically during persistent infection, have been suggested as novel biomarkers for TFI diagnostics. We studied serum IgG antibody responses against C. trachomatis TroA, HtrA and MOMP in 79 subfertile women, of whom 28 had laparoscopically proven TFI. We confirmed that the accuracy of CAT in diagnosing TFI is low, whereas TroA IgG and HtrA IgG are more accurate tests in detecting tubal occlusion and pelvic adhesions. However, the sensitivity and negative predictive value (NPV) of TroA IgG and HtrA IgG are still too low to justify their use as a screening test in clinical practice. Individual immunogenetic profiles combined with TroA and HtrA antibody responses might identify women with the highest risk for developing late complications after C. trachomatis infection

Emättimen mikrobiomi terveyden ylläpitäjänä

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Tavallisimmat emättimen mikrobiologiset tutkimukset - mitä, miksi ja milloin?

publishedVersionPeer reviewe

Predicting tubal factor infertility by using markers of humoral and cell-mediated immune response against Chlamydia trachomatis

Problem: The accuracy of Chlamydia trachomatis antibody test in predicting tubal factor infertility (TFI) is limited, and more accurate methods are needed. Cell-mediated immune response (CMI) is crucial in the resolution of pathogen, but it may play an important role in the pathogenesis of C trachomatis-associated tubal damage. We studied whether combining the markers of C trachomatis-induced CMI to humoral immune response improves the accuracy of serology in TFI prediction. Method of study: Our prospective study consists of 258 subfertile women, of whom 22 (8.5%) had TFI. Women with other causes for subfertility served as a reference group. Serum C trachomatis major outer membrane protein (MOMP) and chlamydial heat-shock protein 60 (cHSP60) IgG antibodies were measured by ELISA. CMI was studied by lymphocyte proliferation assay in vitro. Results: Serological markers were more prevalent in women with TFI than in other subfertile women (40.9% vs 12.3% for MOMP IgG and 27.3% vs 10.2% for cHSP60 IgG). The best test combination for TFI was C. trachomatis MOMP and cHSP60 antibody with an accuracy of 90.3%, sensitivity of 22.7% and specificity of 96.6%. Positive post-test probability of this combination was 54.2%, and negative post-test probability was 12.4%. Adding of the markers of CMI did not significantly improve the accuracy of serology in TFI prediction. Conclusion: The accuracy of TFI prediction increases when the combination of C trachomatis MOMP and cHSP60 antibody tests is used. C trachomatis-induced CMI was common in our study population, but the markers of CMI did not predict TFI.Peer reviewe

Predictive Values of Serum Chlamydia trachomatis TroA and HtrA IgG Antibodies as Markers of Persistent Infection in the Detection of Pelvic Adhesions and Tubal Occlusion

Chlamydia trachomatis IgG antibody testing (CAT) has been used as a screening test for tubal factor infertility (TFI), but as the CAT is only a marker of a past exposure to C. trachomatis and not of late sequelae, the positive predictive value (PPV) of the test is low. The persistence of C. trachomatis in the upper genital tract has been suggested as one of the key mechanisms in the development of TFI. Serum antibodies against C. trachomatis TroA and HtrA, proteins expressed specifically during persistent infection, have been suggested as novel biomarkers for TFI diagnostics. We studied serum IgG antibody responses against C. trachomatis TroA, HtrA and MOMP in 79 subfertile women, of whom 28 had laparoscopically proven TFI. We confirmed that the accuracy of CAT in diagnosing TFI is low, whereas TroA IgG and HtrA IgG are more accurate tests in detecting tubal occlusion and pelvic adhesions. However, the sensitivity and negative predictive value (NPV) of TroA IgG and HtrA IgG are still too low to justify their use as a screening test in clinical practice. Individual immunogenetic profiles combined with TroA and HtrA antibody responses might identify women with the highest risk for developing late complications after C. trachomatis infection.Peer reviewe

Tavallisimmat emättimen mikrobiologiset tutkimukset - mitä, miksi ja milloin?

Vertaisarvioitu.Peer reviewe

Chlamydia trachomatis and Reproductive health

Chlamydia trachomatis infection is the most common bacterial sexually transmitted infection (STI) worldwide. It has been linked to severe reproductive morbidity, including tubal factor infertility (TFI), ectopic pregnancy (EP), miscarriage and preterm delivery (PTD), but the evidence has mostly been based on clinical case-control studies, and the data retrieved from population-based studies have been limited. The aim of this thesis was to study the role of C. trachomatis infection in adverse pregnancy outcomes and subfertility. To evaluate the link between C. trachomatis infection and EP, miscarriage and PTD, we performed a population-based, biobank health registry study. We used national health registries to identify the cases with adverse pregnancy outcomes (n=2950). The cases were linked to the Finnish Maternity Cohort serum bank to obtain samples for serological analysis. C. trachomatis major outer membrane protein (MOMP)–specific IgG antibodies were determined from the serum samples. Our results confirmed the association between C. trachomatis infection and EP, alhough C. trachomatis seroprevalence was lower than expected. The seroprevalence rate and the link between antichlamydial antibodies and EP were strongest among women over 35 years of age. We did not find a serological link between C. trachomatis infection and miscarriage or PTD. Cell-mediated immune response is crucial in the resolution of C. trachomatis infection, but it may play an important role in the pathogenesis of Fallopian tube damage. The link between serum C. trachomatis IgG antibodies and TFI has been well established, and chlamydia antibody testing (CAT) has been introduced as a screening test for TFI in the initial infertility workup to select high-risk patients for further tubal evaluation. We studied the role of C. trachomatis infection in subfertility by measuring C. trachomatis–specific immune responses in a cohort of subfertile women (n=258). Cell-mediated immune response was analyzed from peripheral blood samples by an in vitro lymphocyte proliferation test using C. trachomatis elementary body (EB) and recombinant chlamydial 60 kDA heat shock protein (cHSP60) as lymphocyte-stimulating antigens. Serum C. trachomatis–specific IgG antibody responses were studied using C. trachomatis MOMP, cHSP60, and C. trachomatis TroA and HtrA as antigens. Our aim was to develop a specific and sensitive non-invasive test for TFI prediction in subfertile women to simplify infertility workup. According to our results, the accuracy of C. trachomatis serology in evaluating TFI can be improved by combining serum C. trachomatis MOMP and cHSP60 IgG antibody tests or combining markers of C. trachomatis–specific cell-mediated and humoral immune responses. C. trachomatis may impair fertility by mechanisms other than occluding the Fallopian tubes, such as causing functional tubal damage or inflammation in the endometrium. We studied the role of C. trachomatis infection in unexplained infertility by analyzing C. trachomatis–specific immune responses in women with no specific etiology for subfertility. Neither humoral nor cell-mediated immune response to C. trachomatis were associated with unexplained infertility. The presence of serum antichlamydial IgG antibodies was linked to a prolonged time for spontaneous pregnancy, but pregnancy outcomes, including live birth rate, did not differ between seropositive and seronegative women. The highest prevalence of C. trachomatis infection is seen in young, sexually active women aged 20–25 years. Studies estimating the risk of long-term sequelae following C. trachomatis infection are important, because women diagnosed with chlamydial infection are usually worried and need counseling for their future fertility. Our study, together with other population-based data, suggests that the long-term risks following chlamydial infection are lower than previously thought. Since the development of TFI is multifactorial, C. trachomatis immune markers in TFI prediction are only of modest value. TFI prevalence was low in our study (8.5%), which is in line with the declining trend of C. trachomatis–associated reproductive problems in the population. This results most likely from the successful C. trachomatis screening programs, which have led to the effective treatment of chlamydial infections before they ascend to the upper genital tract. The risk of reproductive problems is, however, significantly higher after recurrent chlamydial infection, and preventive strategies should be planned to recognize the core group at the highest risk for recurrent infection.Chlamydia trachomatiksen aiheuttama klamydia on maailman yleisin bakteerin aiheuttama sukupuolitauti ja raportoidut infektiot ovat lisääntyneet viimeisen kymmenen vuoden aikana. Klamydiainfektio on liitetty hankaliin lisääntymisterveyden ongelmiin, kuten munanjohdinperäiseen lapsettomuuteen, kohdun ulkopuoliseen raskauteen, keskenmenohin ja ennenaikaiseen synnytykseen. Aiemmat tutkimustulokset ovat kuitenkin pohjautuneet tapaus-verrokkitutkimuksiin ja väestötason tutkimuksia on ollut vähän. Tämän väitöstutkimuksen tavoitteena oli tutkia klamydian syyosuutta naisen lisääntymisterveyden ongelmissa. Ensimmäisessä osatyössä tutkimme sairastetun klamydiainfektion yhteyttä kohdun ulkopuoliseen raskauteen, keskenmenoon ja ennenaikaiseen synnytykseen. Tapaukset (n=2950) kerättiin Terveyden ja hyvinvoinnin laitoksen (THL) ylläpitämistä kansallisista rekistereistä ja yhdistettiin Äitiysneuvolaseerumipankkiin (Finnish Maternity Cohort, FMC). Seeruminäytteistä määritettiin IgG vasta-aineita C. trachomatiksen major outer membrane proteiinia (MOMP) vastaan. Tutkimuksemme vahvisti klamydian ja kohdun ulkopuolisen raskauden yhteyden, vaikka C. trachomatis vasta-aineiden esiintyvyys olikin pienempi, kuin mitä aikaisemmissa tutkimuksissa on todettu. Vasta-aineiden esiintyvyys nousi naisen iän mukana, ollen korkein yli 35-vuotiaiden naisten ryhmässä. Klamydiainfektion ja keskenmenojen tai ennenaikaisen synnytyksen välillä emme todenneet yhteyttä. Soluvälitteinen immuunivaste on tärkeä klamydiainfektion paranemisessa, mutta sen tiedetään olevan osallisena kudosvaurioiden kehittymisessä. Seerumin C. trachomatis vasta-aineet ovat yleisiä munanjohdinperäistä lapsettomuutta sairastavilla naisilla ja vasta-aineiden mittaamista onkin esitetty osaksi lapsettomuuden alkututkimuksia, jotta voitaisiin varhaisessa vaiheessa suunnitella kullekin parille sopiva hoito ja nopeuttaa diagnostiikkaa. Tutkimme klamydiainfektion herättämää solu- ja vasta-ainevälitteistä immuunivastetta prospektiivisessa kohorttitutkimuksessa lapsettomuudesta kärsivillä naisilla (n=258). Soluvälitteistä immuunivastetta tutkittiin stimuloimalla perifeerisen veren lymfosyyttejä soluviljelmässä C. trachomatiksen elementary body (EB)- ja klamydian 60kDa heat shock-proteiini (cHSP60) -spesifisillä antigeeneillä. Seerumista määritettiin IgG vasta-aineita C. trachomatiksen MOMP:a ja cHSP60:a vastaan. Tutkimme myös kroonisessa klamydiainfektiossa esiintyvien proteiinien, TroA ja HtrA, aiheuttamaa vasta-ainevälitteistä immuunivastetta. Tavoitteenamme oli kehittää lapsettomuuden alkututkimuksiin soveltuva testi munanjohdinvaurion osoittamiseksi hyödyntämällä klamydiainfektion aiheuttaman immuunivasteen markkereita. Paras tarkkuus saatiin yhdistämällä C. trachomatis MOMP ja cHSP60 vasta-ainetestit, tai yhdistämällä C. trachomatis-spesifinen vasta-aine- ja soluvälitteinen immuunivaste. Sairastettu klamydiainfektio voi heikentää hedelmällisyyttä myös muilla mekanismeilla kuin arpeuttamalla munanjohtimet. Analysoimme klamydiaspesifistä immuunivastetta naisilla, joilla ei löytynyt ilmeistä syytä lapsettomuudelle. Totesimme, että sairastetulla klamydiainfektiolla ei ollut merkitystä lapsettomuushoitojen onnistumiseen eikä raskaustuloksiin. Kuitenkin naisilla, joilla seerumin C. trachomatis IgG vasta-aineet olivat positiiviset, aika spontaaniin raskauteen oli pidempi kuin naisilla, joilla ei ollut vasta-aineita. Tämä väitöstutkimus antaa tärkeää tietoa sairastetun klamydiainfektion merkityksestä naisen koko lisääntymisterveydelle. Suurin osa tartunnoista todetaan nuorilla, 20-25-vuotiailla naisilla, jotka ovat usein huolissaan infektion vaikutuksesta myöhempään lisääntymisterveyteen. Tutkimustuloksemme tukevat muita, hiljattain julkaistuja väestötason tuloksia, joiden mukaan klamydiainfektion vaikutus myöhempään lisääntymisterveyteen onkin vähäisempi kuin mitä aikaisemmissa tutkimuksissa on osoitettu. Munanjohtimien kudosvaurion syntyminen on monitekijäinen prosessi, jonka vuoksi C. trachomatikselle spesifiset immunologiset markkerit eivät pysty ennustamaan tarkasti munanjohdinperäistä lapsettomuutta. Munanjohdinperäisen lapsettomuuden esiintyvyys oli tutkimuskohortissamme matala (8.5%), mikä tukee klamydian aiheuttamien lisääntymisterveyden ongelmien vähenemistä. Klamydian osuus sisäsynnytintulehduksen taustalla on vähentynyt, koska aktiivisen opportunistisen seulonnan ansiosta klamydia diagnosoidaan ja hoidetaan varhaisessa vaiheessa. Lisääntymisterveyden ongelmien riski kuitenkin kasvaa merkittävästi toistuvien klamydiainfektioiden seurauksena, jonka vuoksi terveysvalistus tulisi kohdistaa potilaisiin joilla toistuvia infektioita esiintyy