Waddlia chondrophila, a potential agent of human fetal death.We investigated the zoonotic potential zoonotic potential n. The potential for animal infections to be transmissible to humans. of Waddlia chondrophila, a new Chlamydia-like abortigenic agent in ruminants. Anti-Waddlia antibody reactivity was tested by immunofluorescence Immunofluorescence A technique that uses a fluorochrome to indicate the occurrence of a specific antigen-antibody reaction. The fluorochrome labels either an antigen or an antibody. and Western blot Western blot A technique developed in 1979 that is used to confirm ELISA results. HIV antigen is purified by electrophoresis and attached by blotting to a nylon or nitrocellulose filter. . Waddlia seroprevalence seroprevalence Immunology The proportion of a population that is seropositive–ie, has been exposed to a particular pathogen or immunogen; the seropositivity of a population is calculated as the number of individuals who produce a particular antibody divided was higher in women who had had sporadic and recurrent miscarriages than in control women (p<0.001). Waddlia ssp. may represent a cause of human fetal loss. ********** Approximately 15% of pregnancies end in miscarriage (1). However, a cause is identified in only 50% of these cases. Obligate obligate /ob·li·gate/ (ob´li-gat) pertaining to or characterized by the ability to survive only in a particular environment or to assume only a particular role, as an obligate anaerobe. intracellular bacteria, which fail to grow on media used routinely to isolate human pathogens, could represent yet unrecognized agents of miscarriage. Chlamydia trachomatis Chlamydia tra·cho·ma·tis n. A species of Chlamydia that causes trachoma, inclusion conjunctivitis, lymphogranuloma venereum, nonspecific urethritis, and proctitis in humans. , an intracellular bacterium, is the world's most common sexually transmitted bacterial pathogen (2). Because C. trachomatis Noun 1. C. trachomatis - bacteria responsible for the sexually transmitted diseases chlamydia and lymphogranuloma venereum Chlamydia trachomatis chlamydia - coccoid rickettsia infesting birds and mammals; cause infections of eyes and lungs and genitourinary is asymptomatic, most infected women remain untreated. Although the effect of C. trachomatis infection on pregnancy outcome is unclear, an increased prevalence of C. trachomatis immunoglobulin (Ig) G antibodies in women with a history of miscarriage has been observed (2,3). Chlamydophila abortus Chlamydophila abortus is a species in Chlamydiae that causes abortion and fetal death in mammals, including humans. Chlamydophila abortus was previously classified as Chlamydia psittaci along with all Chlamydiae except Chlamydia trachomatis. is the most common etiology of abortion in ruminants (4) and can also cause miscarriage in pregnant women exposed to infected animals. Waddlia chondrophila, another Chlamydiales, is a new abortigenic agent in bovines (5,6). This obligate intracellular bacterium was isolated from aborted fetuses in the United States United States, officially United States of America, republic (2005 est. pop. 295,734,000), 3,539,227 sq mi (9,166,598 sq km), North America. The United States is the world's third largest country in population and the fourth largest country in area. (7) and in Germany (8). A serologic se·rol·o·gy n. pl. se·rol·o·gies 1. The science that deals with the properties and reactions of serums, especially blood serum. 2. study further supported the abortigenic role of Waddlia in bovine species (6). Moreover, infection of bovine fetuses with Waddlia was associated with their deaths within 2 weeks (6). In this study, we tested women with sporadic and recurrent miscarriages for antibody reactivity against Waddlia and compared seroprevalence with that found in a control group of women with uneventful term pregnancies. The Study From July 2004 to March 2005, we studied 69 women with sporadic miscarriages (SM), 200 women who had suffered from recurrent miscarriages (RM), and 169 control women who had had uneventful pregnancies (Table 1). The RM group comprised women who had [greater than or equal to] 3 miscarriages (1) and who had attended the Recurrent Miscarriage Clinic of St Mary's Hospital, London, the largest specialist referral center in Europe. Immunofluorescence tests were performed (9); we used W. chondrophila strain ATCC ATCC American Type Culture Collection, see there VR-1470 as antigen and we screened sera at a 1:64 dilution with FluolineH (bioMerieux, Marcy l'Etoile, France). Antigen was isolated as described (10,11). Mice and rabbit anti-Waddlia antibodies were used as positive controls with a fluorescein-conjugated anti-mouse and anti-rabbit globulin globulin, any of a large family of proteins of a spherical or globular shape that are widely distributed throughout the plant and animal kingdoms. Many of them have been prepared in pure crystalline form. . Sera that exhibited an Ig titer titer /ti·ter/ (ti´ter) the quantity of a substance required to react with or to correspond to a given amount of another substance. [greater than or equal to] 64 were tested for IgG and IgM reactivity by using corresponding anti-human Ig fluorescein fluorescein /flu·o·res·ce·in/ (fldbobr-res´en) a fluorescing dye; its sodium salt is used as a tracer in retinal angiography and as a diagnostic aid for revealing corneal trauma and fitting contact lenses. (FluolineG or FluolineM, bioMerieux) and serial 2-fold dilutions of serum. IgG and IgM positivity cut-offs were [greater than or equal to] 1:64 and [greater than or equal to] 1:32, as proposed for other chlamydia-like organisms (4). One hundred women had an anti- Waddlia IgG titer [greater than or equal to] 64 (Table 1). Seroprevalence was higher for patients who had sporadic (31.9%) and recurrent (33%) miscarriages than that for women who had had uneventful pregnancies (7.1%, p<0.001 when comparing either SM or RM groups to controls). One woman had a positive lgM titer of 64 and an IgG titer of 512. To confirm the specificity of immunofluorescence, we performed Western blot analyses on all Waddlia-positive sera samples. Western blot was performed (9), but Waddlia was used as the antigen. A polyclonal polyclonal /poly·clo·nal/ (-klon´'l) 1. derived from different cells. 2. pertaining to several clones. polyclonal derived from different cells; pertaining to several clones. peroxidase-labeled anti-human IgG (Dako, Glostrup, Denmark) was used as a secondary antibody A secondary antibody is an antibody that binds to primary antibodies or antibody fragments. They are typically labeled with probes that make them useful for detection, purification or cell sorting applications. . The presence of anti-Waddlia IgG antibodies was confirmed by Western blot in 97 of the 100 positive samples by immunofluorescence (Table 1; Figure 1, panels B and C). By using Waddlia spp. hyperimmune hyperimmune /hy·per·im·mune/ (hi?per-i-mun´) possessing very large quantities of specific antibodies in the serum. hyperimmune possessing very large quantities of specific antibodies in the serum. mouse and rabbit sera and corresponding peroxidase-conjugated anti-mouse/rabbit sera, we obtained similar patterns of 61-, 55-, 53-, 45-, 41-, 38- and 30-kDa proteins (Figure 1, panels D and E). Antibody reactivity against the 55-, 53-, 45-, 41- and 38-kDa proteins disappeared after adsorption adsorption, adhesion of the molecules of liquids, gases, and dissolved substances to the surfaces of solids, as opposed to absorption, in which the molecules actually enter the absorbing medium (see adhesion and cohesion). with 10 (8) Waddlia antigen for 48 hours, which demonstrated the specificity of the antibody response (Figure 1, panel F). [FIGURE 1 OMITTED] For further statistical analyses, only patients whose samples were positive for Waddlia spp. by immunofluorescence and confirmed by Western blot were considered seropositive seropositive /se·ro·pos·i·tive/ (-poz´i-tiv) showing positive results on serological examination; showing a high level of antibody. se·ro·pos·i·tive adj. for Waddlia spp. (n = 97). In all age groups, the rate of Waddlia seropositivity Seropositivity is the presence of a certain antibody in a blood sample. A patient with seropositivity for a particular antigen or agent is termed seropositive. was higher in patients who had miscarried than in those who had not (data not shown). Moreover, most women with anti-Waddlia antibodies did not exhibit serologic reactivity against C. trachomatis (Table 2). In a multivariate logistic regression In statistics, logistic regression is a regression model for binomially distributed response/dependent variables. It is useful for modeling the probability of an event occurring as a function of other factors. adjusted for age, ethnicity, contact with animals and C. trachomatis serostatus (Figure 2), miscarriage (SM/RM) remained strongly associated with Waddlia seropositivity (odds ratio [OR] 4.9, 95% confidence interval confidence interval, n a statistical device used to determine the range within which an acceptable datum would fall. Confidence intervals are usually expressed in percentages, typically 95% or 99%. [CI] 2.5-9.4). In this model, miscarriage was also independently associated with age (OR 2.9, 95% CI 2.0-4.1) and C. trachomatis seropositivity (OR 2.3, 95% CI 1.2-4.5). Additional multivariate models confirmed the association between Waddlia IgG seropositivity and miscarriage, with ORs ranging from 4.9 to 6.2. [FIGURE 2 OMITTED] Noninfectious causes of miscarriage have also been recorded (Table 2). When studying only the 322 patients without known concurrent conditions, the strong correlation between Waddlia seropositivity and miscarriage was still present (68/195 [34.8%] of patients who had had a miscarriage vs. 10/127 [7.9%] of patients who had not miscarried had anti- Waddlia antibodies; p<0.001). Waddlia seropositivity was associated with early miscarriage (p<0.001, Table 2). No difference in ethnicity was observed between those who were IgG positive for Waddlia spp. and those who were negative. Moreover, there was no difference in C. trachomatis, Chlamydophila pneumoniae, or Cp. psittaci seropositivity between women who had anti-Waddlia antibodies and those who did not, which suggests that Waddlia antibodies do not cross-react with Chlamydiaceae. Moreover, only 7 (1.6%) of 438 patients had a Parachlamydia IgG titer [greater than or equal to] 64. With such a low Parachlamydia prevalence, cross-reactivity with this chlamydia-like organism is unlikely to explain the high Waddlia seroprevalence observed in the miscarriage groups. Women who were IgG seropositive for Waddlia spp. were more likely to have had contact with animals. In a multivariate logistic regression model adjusted for age, ethnicity, and C. trachomatis serostatus, those who had had previous contact with animals were more likely to exhibit anti-Waddlia antibodies (OR 1.7, 95% CI 1.0-2.9, Figure 2). In this model, Waddlia IgG-positive serologic test serologic test Lab medicine A test that measures components–eg, antibodies, complement, and reactions–eg, complement fixation, agglutination, precipitation, etc, that reflect immune status, especially antibody titers. Cf Seroconversion. results were also independently associated with age (OR 2.1, 95% CI 1.4-3.1). Conclusions This study demonstrates a strong association between the presence of W. chondrophila-specific IgG antibodies and early fetal loss. Cross-reactivity with other microorganisms seems an unlikely explanation for our results because W. chondrophila did not react with monoclonal or polyclonal antisera directed against Rickettsia rickettsia (rĭkĕt`sēə), any of a group of very small microorganisms, many disease-causing, that live in vertebrates and are transmitted by bloodsucking parasitic arthropods such as fleas, lice (see louse), and ticks. , Coxiella, Wolbachia, Anaplasma, and Chlamydia chlamydia (kləmĭd`ēə), genus of microorganisms that cause a variety of diseases in humans and other animals. Psittacosis, or parrot fever, caused by the species Chlamydia psittaci, spp. (8,12). We did not detect any cross-reactivity of W. chondrophila with C. trachomatis, Cp. pneumoniae, and Cp. psittaci. Moreover, the molecular weights of Waddlial immunoreactive immunoreactive exhibiting immunoreactivity. proteins obtained by Western blot are clearly different than those reported for C. trachomatis or Cp. pneumoniae (13). With the exception of 1 patient who had IgM, only IgG antibody reactivity against W. chondrophila was observed. Because IgG antibodies may persist for years after an acute infection has resolved (3), the underlying mechanism of miscarriage due to W. chondrophila may involve reactivation reactivation to become active after a period of quiescence or, as in bacterial and viral infections, latency. cross reactivation of a latent asymptomatic waddlial infection, endometrial endometrial /en·do·me·tri·al/ (en?do-me´tre-il) pertaining to the endometrium. endometrial, n relating to the end-ometrium or cavity of the uterus. damage from a past waddlial infection, or an immune response immune response n. An integrated bodily response to an antigen, especially one mediated by lymphocytes and involving recognition of antigens by specific antibodies or previously sensitized lymphocytes. to an epitope epitope: see immunity. shared by a waddlial and fetal antigen, as proposed for C. trachomatis (3). The association we found between contact with animals and positive serologic results for Waddlia spp. raises the zoonotic potential of this bacterium. This hypothesis is further supported by the range of hosts for Waddliaceae (4). Other modes of transmission are possible (e.g., contaminated contaminated, v 1. made radioactive by the addition of small quantities of radioactive material. 2. made contaminated by adding infective or radiographic materials. 3. an infective surface or object. water) because free-living amebae may serve as hosts for Waddlia spp. (14) and are widespread in water networks (15). Waddlia spp. may also be transmitted through ingestion ingestion /in·ges·tion/ (-chun) the taking of food, drugs, etc., into the body by mouth. in·ges·tion n. 1. The act of taking food and drink into the body by the mouth. 2. of contaminated cow milk. Finally, Waddlia spp. might represent a sexually transmitted disease sexually transmitted disease (STD) or venereal disease, term for infections acquired mainly through sexual contact. Five diseases were traditionally known as venereal diseases: gonorrhea, syphilis, and the less common granuloma inguinale, . Further investigations are urgently needed to define how Waddlia spp. infection may be acquired. To confirm the role of W. chondrophila in miscarriage, it will be important to isolate this intracellular bacterium from miscarriage products or confirm its presence in the placenta placenta (pləsĕn`tə) or afterbirth, organ that develops in the uterus during pregnancy. It is a unique characteristic of the higher (or placental) mammals. In humans it is a thick mass, about 7 in. by immunohistochemistry or PCR PCR polymerase chain reaction. PCR abbr. polymerase chain reaction Polymerase chain reaction (PCR) . This may be difficult to achieve, if, as suggested for C. trachomatis (3), W. chondrophila causes miscarriage indirectly, e.g., through increased cytokine Cytokine Any of a group of soluble proteins that are released by a cell to send messages which are delivered to the same cell (autocrine), an adjacent cell (paracrine), or a distant cell (endocrine). production or molecular mimicry molecular mimicry Immunology A mechanism that may explain some forms of autoimmune disease, where the immune system attacks self antigens that are structurally similar to nonself antigens with fetal antigens. To our knowledge, this work provides the first evidence that W. chondrophila may be implicated im·pli·cate tr.v. im·pli·cat·ed, im·pli·cat·ing, im·pli·cates 1. To involve or connect intimately or incriminatingly: evidence that implicates others in the plot. 2. in human fetal death. Acknowledgments We thank all midwives and doctors who participated in this study. Their involvement was essential to the whole process, and they enthusiastically gave their time to provide information and samples. We thank Andre Baud for computer assistance, Ludivine Pugin for technical help, and Philip E. Tarr for critical review of the manuscript. The research of the group directed by G.G. is supported by grants from the Swiss National Science Foundation The Swiss National Science Foundation is a science research support organization mandated by the Swiss Federal Government. The SNSF was established in 1952 as a foundation under private law. Its secretariat is based in Berne. (FN3200BO105885 and FN3200BO-116445) and COST Action 855 (European Cooperation in the field of Scientific and Technical Research). References (1.) Rai R, Regan L. Recurrent miscarriage. Lancet. 2006;368:601-11. (2.) Mardh PA. Influence of infection with Chlamydia trachomatis on pregnancy outcome, infant health and life-long sequelae sequelae Clinical medicine The consequences of a particular condition or therapeutic intervention in infected offspring. Best Pract Res Clin Obstet Gynaecol. 2002;16:847-64. (3.) Witkin SS, Ledger WJ. Antibodies to Chlamydia trachomatis in sera of women with recurrent spontaneous abortions. Am J Obstet Gynecol. 1992;167:135-9. (4.) Corsaro D, Greub G. Pathogenic potential of novel chlamydiae Chlamydiae is a bacterial phylum whose members are obligate intracellular pathogens.[1] Many Chlamydiae coexist in an asymptomatic state within specific hosts, and it is widely believed that these hosts provide a natural reservoir for these species. and diagnostic approaches to infections due to these obligate intracellular bacteria. Clin Microbiol Rev. 2006;19:283-97. (5.) Rurangirwa FR, Dilbeck PM, Crawford TB, McGuire TC, McElwain TF. Analysis of the 16S rRNA gene of micro-organism WSU WSU Washington State University WSU Wayne State University WSU Wichita State University WSU Wright State University WSU Weber State University WSU Western State University College of Law WSU Winona State University WSU Walter Sisulu University 86-1044 from an aborted bovine foetus reveals that it is a member of the order Chlamydiales: proposal of Waddliaceae fam. nov., Waddlia chondrophila gem nov., sp. nov. Int J Syst Bacteriol. 1999;49: 577-81. (6.) Dilbeck-Robertson P, McAllister MM, Bradway D, Evermann JF. Results of a new serologic test suggest an association of Waddlia chondrophila with bovine abortion. J Vet Diagn Invest. 2003;15: 568-9. (7.) Dilbeck PM, Evermann JF, Crawford TB, Ward AC, Leathers CW, Holland CJ, et al. Isolation of a previously undescribed rickettsia from an aborted bovine fetus. J Clin Microbioil. 1990;28:814-6. (8.) Henning K, Schares G, Granzow H, Polster U, Hartmann M, Hotzel H, et al. Neospora caninum and Waddlia chondrophila strain 2032/99 in a septic stillborn stillborn /still·born/ (-born) born dead. still·born adj. Dead at birth. stillborn, n an infant who is born dead. stillborn born dead. calf. Vet Microbiol. 2002;85:285-92. (9.) Greub G, Boyadjiev I. La Scola B, Raoult D, Martin C. Serological serological pertaining to or emanating from serology. serological test one involving examination of blood serum usually for antibody. hint suggesting that Parachlamydiaceae are agents of pneumonia in polytraumatized intensive care patients. Ann N Y Acad Sci. 2003;990:311-9. (10.) Greub G, Raoult D. Crescent bodies of Parachlamydia acanthamoeba Acanthamoeba /Acan·tha·moe·ba/ (ah-kan?thah-me´bah) a genus of free-living ameboid protozoa (order Amoebida) found usually in fresh water or moist soil. Certain species, such as A. astronyxis, A. castellanii, A. culbertsoni, A. and its life cycle within Acanthamoeba polyphaga: an electron micrograph study. Appl Environ Microbiol. 2002;68:3076-84. (11.) Greub G, Mege JL, Raoult D. Parachlamydia acanthamoebae enters and multiplies within human macrophages Macrophages White blood cells whose job is to destroy invading microorganisms. Listeria monocytogenes avoids being killed and can multiply within the macrophage. and induces their apoptosis. Infect Immun. 2003;71:5979-85. (12.) Kocan KM, Crawford TB, Dilbeck PM, Evermann JF, McGuire TC. Development of a rickettsia isolated from an aborted bovine fetus. J Bacteriol. 1990;172:5949 55. (13.) Biendo M, Eb F, Lefebvre JF, Orfila J. Limits of the microimmunofluorescence test and advantages of immnnoblotting in the diagnosis of chlamydiosis. Clin Diagn Lab Immunol. 1996;3:706-9. (14.) Michel R, Steinert M, Zoller L, Hauroder B, Henning K. Free-living Amoebae may serve as hosts for the Chlamydia-like bacterium Waddlia chondrophila isolated from aborted bovine foetus. Acta Protozoologia. 2004;43:37-42. (15.) Thomas V, Herrera-Rimann K, Blanc DS, Greub G. Biodiversity of amoebae and amoeba-resisting bacteria in a hospital water network. Appl Environ Microbiol. 2006;72:2428-38. Address for correspondence: Gilbert Greub, Center for Research on Intracellular Bacteria, Institute of Microbiology, University Hospital Center and University of Lausanne The University of Lausanne (in French: Université de Lausanne) or UNIL in Lausanne, Switzerland was founded in 1537 as a school of theology, before being made a university in 1890. Today about 10,000 students and 2200 researchers study and work at the university. , Bugnon 48, 1011 Lausanne, Switzerland; email: gilbert.greub@chuv.ch Dr Baud is a physician working in the Department of Obstetrics and Gynecology obstetrics and gynecology Medical and surgical specialty concerned with the management of pregnancy and childbirth and with the health of the female reproductive system. of Professor Lesley Regan (London) and in the group of Gilbert Greub (Lausanne). His current research focuses on emerging infectious causes of adverse pregnancy outcomes. David Baud,*[dagger] Vincent Thomas,* Aliaa Arafa, [dagger] Lesley Regan, [dagger] and Gilbert Greub* *University of Lausanne, Lausanne, Switzerland; [dagger] St Mary's Hospital, London, UK
Table 1. Characteristics of study patients according to
miscarriage history *
Controls Sporadic
(n = 169), miscarriages
no. (%) (n = 69), no. (%,
Characteristics p value [dagger])
Age, y
Median (IQR, p value 30.4 (25-35) 31.7 (27-36, 0.18)
[dagger])
No. pregnancies
1 90 (53.3) 32 (46.4)
2 46 (53.3) 19 (27.5, 0.67
[double dagger])
>2 33 (19.5) 18 (26.1)
Mean (SD, p valuet) 1.8 (1.1) 2.1 (1.5, 0.23)
Parity
0 0 (0) 49 (71)
1 115 (68.1) 11 (15.9, <0.001
[double dagger])
2 31 (18.3) 4 (5.8)
>2 23 (13.6) 5 (7.3)
Mean (SD, p value 1.54 (0.95) 0.52 (1.01, <0.001)
[dagger])
Miscarriages
Early ([less than or 0 52 (75.4, <0.001)
equal to] 12 weeks)
Late (>12 weeks) 0 13 (18.9, <0.001)
Stillbirth (>24 weeks) 0 4 (5.8, <0.001)
Alive child 169 (100) 21 (30.4, <0.001)
Ethnicity
White 80 (47.3) 34 (49.3, 0.79)
Black 35 (20.7) 10 (14.5, 0.27)
Asian 31 (18.3) 18 (26.1, 0.18)
Other 31 (18.3) 6 (8.7, 0.35)
Contact with animals 22 (13) 11 (15.9, 0.82)
Cat 18 (10.7) 5 (7.3, 0.42)
Dog 15 (8.9) 6 (8.7, 0.97)
Fish 1 (0.6) 0 (0, 1)
Rodents 0 2 (2.9, 0.08)
Other 3 (1.8) 0 (0, 0.63)
Chlamydia trachomatis 15 (8.9) 9 (13, 0.33)
(IgG titer>50)
Positive serology for
Waddlia spp.
IgG titer >64 12 (7.1) 22 (31.9, <0.001)
IgG titer >128 6 (3.6) 10 (14.5, 0.002)
Western blot Waddlia
IgG
[less than or equal 12 (7.1) 22 (31.9, <0.001)
to]1 band
[less than or equal 6 (3.6) 15 (21.7, <0.001)
to] 2 bands
[less than or equal 3 (1.8) 9 (13, <0.001)
to] 3 bands
[less than or equal 1 (0.6) 9 (13, <0.001)
to] 2 specific
bands [section]
Recurrent
miscarriages
(n = 200), no. (%
Characteristics p value [dagger])
Age, y
Median (IQR, p value 35.4 (31-39, <0.001)
[dagger])
No. pregnancies
1 0
2 0(0, <0.001
[double dagger])
>2 200 (100)
Mean (SD, p valuet) 5.2 (3, <0.001)
Parity
0 113 (56.5)
1 59 (29.5, <0.001
[double dagger])
2 19 (9.5)
>2 9 (4.5)
Mean (SD, p value 0.64 (0.9, <0.001)
[dagger])
Miscarriages
Early ([less than or 196 (98, <0.001)
equal to] 12 weeks)
Late (>12 weeks) 51 (25.5, <0.001)
Stillbirth (>24 weeks) 11 (5.5, <0.002)
Alive child 78 (39, <0.001)
Ethnicity
White 132 (66, <0.001)
Black 22 (11, 0.01)
Asian 34 (17, 0.74)
Other 8(4, 0.002)
Contact with animals 70 (35, <0.001)
Cat 37 (18.5, 0.035)
Dog 28 (14, 0.13)
Fish 8 (4, 0.043)
Rodents 7 (3.5, 0.017)
Other 12 (6, 0.041)
Chlamydia trachomatis 39 (19.5, 0.004)
(IgG titer>50)
Positive serology for
Waddlia spp.
IgG titer >64 66 (33, <0.001)
IgG titer >128 29 (14.5, <0.001)
Western blot Waddlia
IgG
[less than or equal 63 (31.5, <0.001)
to]1 band
[less than or equal 51 (25.5, <0.001)
to] 2 bands
[less than or equal 28 (14, <0.001)
to] 3 bands
[less than or equal 22 (11, <0.001)
to] 2 specific
bands [section]
* Controls, women with uneventful pregnancies, IQR, interquartile
range.
[dagger] Compared with control group.
[double dagger] p value for ordered categories (Pearson [chi square]
test).
[section] According to Waddlia hyperimmune mouse serum, bands at 61-,
55-, 53-, 45-, 41-, 38-, and 30-kDa are considered specific for
Waddlia spp.
Table 2. Characteristics of patients in the study according
to their Waddlia serostatus
Waddlia Waddlia p value
negative * positive
(n = 341) [dagger]
(n = 97)
Age, y
Median (interquartile 33 (28-37) 36 (31-39) <0.001
range)
No. pregnancies
1 107 (31.4) 15 (15.5)
2 62 (18.2) 16 (16.5) 0.006 [double
dagger]
>2 172 (50.4) 66 (68)
Mean (SD) 3.2 (2.6) 4 (3.1) 0.005
Parity
0 108 (31.7) 54 (55.7)
1 156 (45.8) 29 (29.9) 0.001 [double
2 49 (14.4) 5 (5.2) dagger]
2 28 (8.2) 9 (9.3)
Mean (SD) 1.04 (l.04) 0.7 (1) <0.001
Miscarriages
Early ([less than or 170 (49.9 81 (83.5) <0.001
equal to] 12 wk)
Late (>12 wk) 47 (13.8) 17 (17.5) 0.36
Stillbirth (>24 wk) 12 (3.5) 3 (3.1) 1
Alive child 228 (66.9) 40 (41.2) <0.001
Ethnicity
White 184 (54) 62 (63.9) 0.08
Black 52 (15.3) 15 (15.5) 0.96
Asian 70 (20.5) 13 (13.4) 0.11
Other 29 (8.5) 7 (7.2) 0.68
Contact with animals 76 (22.3) 34 (35.1) 0.011
Cat 42 (12.3) 18 (18.6) 0.12
Dog 35 (10.3) 14 (14.4) 0.25
Fish 6 (1.8) 3 (3.1) 0.42
Rodent 7 (2.1) 2 (2.1) 1
Other 12 (3.5) 3 (3.1) 1
Noninfectious miscar- 97 (28.5) 19 (19.6) 0.08
riage causes
Autoimmune disease 19 (5.6) 1 (1) 0.09
Hypertensive disorder 8 (2.4) 3 (3.1) 0.71
Endocrine pathology 13 (3.8) 2 (2.1) 0.54
Anatomic abnormalities 4 (1.2) 2 (2.1) 0.62
Additional serologies
(titers)
Chlamydia trachomatis 49 (14.7) 14 (14.3) 0.98
(IgG [greater than or
equal to] 50)
Clamydophila pneumo- 161 (47.2) 38 (39.2) 0.16
niae (IgG [greater
than or equal to] 64)
Cp. psittaci (IgG 20 (5.9) 6 (6.2) 1
[greater than or equal
to] 1/64)
* Patients with a Waddlia immunoglobulin G (IgG) titer <64 (n = 338)
or not confirmed by Western blot analysis (n = 3).
[dagger] Patients with a Waddlia IgG titer >64 and confirmed by
Western blot analysis.
[double dagger] p value for ordered categories (Pearson [chi square]
test).
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