THE RATE OF CHLAMYDIA TRACHOMATIS, MYCOPLASMA HOMINIS AND UREAPLASMA UREALY- TICUM IN FEMALES WITH HABITUAL ABORTION AND
ITS COMPARISON WITH CONTROL GROUP
M.H. Salari and N. Badami
Department of Pathobiology, School of Public Health and Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, Iran
Abstract: Females abortion is one of the most important sequela of genital infection with Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum.
In this study frequency of Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum was studied in 125 females with habitual abortion by direct and indirect immunofluorescence tests and culture method and compared with 250 normal population. The results obtained were as follow:
Mycoplasma hominis was isolated from 18 (14.4%) females with habitual abortion and 18 (7.2%) normal population (P=0.0139). Ureaplasma urealyticum was isolated from 39 (31.2%) females with habitual abortion and 48 (19.2%) normal population (P= 0.0045).
Chlamydia trachomatis was detected by direct immunofluorescence test in 9 (7.2%) of cases and 2 (0.8%) of control groups (P=0.0002). The antibody titer against D-K serotypes of Chlamydia trachomatis was also measured. The valuable titer of antibody (>1/16) was detected in 15 (12%) of cases and 8 (3.2%) of control groups (P=0.0004).
The results show that Chlamydia trachomatis and Ureaplasma urealyticum may be responsible for some cases of abortion.
Acta Medica Iranica: 40(2): 79-82; 2002.
Key Words: Chlamydia trachomatis, mycoplasma hominis, ureaplasma urealyticum, females, habitual abortion
INTRODUCTION
Females genital tract is a suitable place for growth of many micro-organisms. Some of these micro-organisms such as Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, Gardnerella vaginalis, Listeria monocytogenes and
Correspondence:
M.H. Salari, Department of Pathobiology, School of Public Health and Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, Iran
P.O.Box: 14155-6446.
Tel: 03526226559
E.mail: Msalari [email protected]
Neisseria gonorrhoeae upon localization and colonization at the appropriate anatomical site may cause various pathological disorders like cervicitis, vaginitis, urethritis, endometritis, salpingitis, bartho- linitis. The pathological disorders may lead to pelvisitis, ectopic pregnancy, abortion and infertility and cervical displasia. It was reported that some of these microorganisms produced noraminidase like substance which showed to induce certain disorders in blastocyste of murine model and might cause a similar pathological disorders in human (1-4).
The purpose of this study was to investigate the rate of Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum in females with habitual abortion and their comparison with the control group.
MATERIALS AND METHODS
In this study, 125 samples from females with habitual abortion referred to Imam Khomeini and Mirza Koochak Khan hospitals of Tehran and 250 samples from healthy volunteers (control group) were collected during 1996-1998.
By using speculum, 4 sterile cotton swabs were used to collect specimens. One of the swabs was transferred to pplo broth medium, the two other were used for gram stain and direct fluorescence test (DIF), the forth swab was kept at - 70°C for further validation. Then the samples were transported to laboratory for further tests. Five ml of blood sample was collected from each case for titration of antichlamydial antibodies. The media containing suspected Mycoplasma and Ureaplasma were passed through membrane filter of 0.22 milipore by pressure, then inoculated in to urea and argenine containing media and incubated at 37°C under 5%
CO2. When the medium color changed, the sample was transferred to pplo agar medium for further confirmation (5-8). Serum samples were checked within 3 different following pooled Chlamydia antigens:
The rate of chlamidydia and … in females with …
80 1- A, B, B1, C,
2- D, E, F, G, H, I, J, K, 3- L1, L2, L3
A serial dilution of each serum sample was prepared and added to different aceton fixed antigens. After 30 minutes incubation in 37°C in moist condition, slides were washed with PBS, for 10 minutes and then dried at room temperature. Then the labeled anti-human fluorescein isothiocyanate conjugate (FITC) was added to each slide and incubated at 37°C in moist condition for 30 minutes.
The slides were washed with PBS and PBS containing evanseblue, and a drop of buffered glycerol solution was added to each of them. Then they were observed under fluorescence microscope (9-12).
RESULTS
In this study, 125 samples from females with habitual abortion and 250 samples from control group (Healthy volunteers) were collected. The samples were processed by culture and serological tests. The results are as follow:
Table 1 shows frequency distribution of females on the basis of age, signs of disease and level of education in case and control groups. In regard of the profession of the subjects under investigation, out of 125 cases, 97 (77.6%) were housewives and 28 (22.4%) were employees. Also in control group, 155 (62%) were housewives and 95(38%) were employees. Detective rate of Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum in
the case group were 7.2%, 14.4% and 31.2% , respectively . These rates in control group were 0.8%, 8.7% and 19.2% respectively (Table 2). The obtained results of antibody titration against D-K serotypes of Chlamydia trachomatis in case (12%) and control groups (3.2%) are shown in Table 3 (P=0.0004).
Z statistical analysis test with 99% confidence, showed that the rates of Chlamydia
trachomatis and Ureaplasma urealyticum in case and control groups had significant differences.
DISCUSSION
This study was undertaken to investigate the rate of Mycoplasma hominis, Ureaplasma urealyticum and Chlamydia trachomatis in involved females with habitual abortion. The obtained results of this study are shown in Tables 2-3 and indicated as follows:
Mycoplasma hominis was isolated from 14.4%
of case and 8.7% of control groups (P=0.0139).
The rate of this bacterium was reported in 14-15%
case and 9-10% in control groups by Maitrayee RM, et al (1994). These results are nearly the same as our findings (13).
Isolation rate of Ureaplasma urealyticum in case and control groups was 31.2% and 19.2%, respectively ( P = 0.0045). The rate of this Microorganism in females with abortion and control group was reported 24-54% and 15-20%, respectively by Nancy EL, et al (1994), Maitrayee RM, et al (1994) and Berg TG, et al (1999) (13-15).
Table 1. Frequency distribution of under study females on the basis of age, signs of disease and level of education Age
( in year )
Case group n = 125
No %
Control group n = 250
N %
<20 3 ( 2.4 ) 13 ( 5.2 )
21-24 37 (29.6) 61 (24.4)
25-29 37 (29.6) 70 ( 28 )
30-34 20 ( 16 ) 59 (23.6)
35-39 24 (19.2) 37 (14.7)
>40 4 ( 3.2 ) 10 ( 4 )
Signs of disease:
Discharge 15 ( 12 ) __
Spotting 54 (43.2) __
Itching and burning 36 ( 28 ) __
Urine frequency 24 (19.2) __
Pain beneath abdomen 26 (36.3) __
Painful intercourse 46 (36.8) __
Painful menstrual cycle 46 (36.8) __
Spotting after sexual 11 ( 8.8) __
STD of sex partner 2 ( 6 ) __
Self STD 5 ( 16 ) __
Level of education:
Illiterate 9 ( 7.2 ) 17 ( 6.8 )
Primary education 27 (21.6) 43 (17.2)
High school education 59 (47.2) 107 (42.8)
Higher education 30 ( 24 ) 83 (33.2)
Acta Medica Iranica, Vol 40, No 2 (2002)
81
Table 2. Frequency distribution of under study females on the basis of isolated bacteria Case group
Bacteria n = 125 No %
Control group n=250
No % Z P
Mycoplasma 18 (14.4) Hominis
18 ( 8.7 ) 2.23 0.0139
Ureaplasma 39 (31.2)
Urealyticum 48 (19.2) 2.41 0.0045
Chlamydia 9 ( 7.2) trachomatis
(Direct-IF)
2 ( 0.8 ) 3.5 0.0002
Table 3. Frequency distribution of under study females on the basis of specific antibody titer against Chlamydia trachomatis
Antibody titer Case group n = 125
1/128 1/64 1/32 1/16
Control group n = 250 1/128 1/64 1/32 1/16
Z P
Number 1 2 4 8 0 2 2 4
Percentage 0.8 1.6 3.2 6.4 0 0.8 0.8 1.6 3.35 0.0004
Chlamydia trachomatis was observed in 7.2% of cases and 0.8% of control groups by Direct-IF method ( P = 0.0002). Also specific antibody (IgG) titer against D-K serotypes of Chlamydia trachomatis was detected in 14.4% of case and 3.2% of control groups (P=0.0004). These results are in accordance with the studies of Amortegul AJ, et al (1986) and Witkins, et al (1992) (11,16).
The detective rates of Ureaplasma urealyticum and Chlamydia trachomatis showed significant differenees between the case and control groups.
Therefore the results show that these bacteria may be responsible for some cases of abortion.
Acknowledgments
Authors wish to thank Dr K. Ghazi Saeedi, Dr M.
Mahmudi, A. Karmostagee, R. Hafezi, G.R Hassanpour, N. Iranparast, F. Amin Harati, N.
Mardani from Pathobiology Department of School of Public Health, Tehran University of Medical Sciences, Dr M.M. Aghsa from Imam Khomeini hospital and also A.A Hanafi-Bojd, M.M. Hakimian and F. Falah from Yazd Center of Education and Health Research.
REFERENCES
1. Waites KB, Robinson DT. Mycoplasma and Ureaplasma. In: Murray PR, Baron EJ, Pfaller MA, Tenover FC, Yolker RH. Manual of clinical microbiology, 7th edition. USA: ASM Press;
1999; 782-94.
2. Schachter J, Stamm WE. Chlamydia. In:
Murray PR, Baron EJ, Pfallor MA, Tenover FC, Yolker RH. Manual of clinical microbiology, 7th edition. USA: ASM Press ; 1999; 795-806.
3. Taylor RD. Infections due to species of Mycoplasma and Ureaplasma: An update. Clin Infect Dis 1996; 23: 671-3.
4. Romana C, Salleras LSM. Latent listeriosis may cause habitual abortion intrauterine death, fetal malformation. Acta Microbiol Hung 1989;
36: 171-172.
5. Berlin OGW, Drew WL, Edelstein MAC, Garcia LS, Roberts GD. Chlamydia, Mycoplasma and Rickettsia. In: Baron EJ, Fingold SM. Bailey and scott,s diagnostic microbiology, 8th edition. USA: Mosby Company; 1990; 558-71.
6. Marmion BP. Mycoplasma, Acholeplasma and Ureaplasma. In: Collee JG, Duguid JP, Fraser AG, Marmion BP. Makie and McCartney Practical medical microbiology, 13th edition, Sangapore: Churchill Livingstone; 1989; 745-79.
7. Tomas JG, Long KS. Chlamydia, Mycoplasma and Ureaplasma species. In: Mahon CR, Manaselis G. Textbook of diagnostic microbiology, 2nd edition. USA: Saunders Company 2000; 635-65.
The rate of chlamidydia and … in females with …
82 8. Witkin SS. Testing for Chlamydia antibodies in recurrent spontaneous abortion. Fertil Steril. 2000;
73: 656-7.
9. Borisov I. The direct immunoflouresence test for the diagnosis of Chlamydia trachomatis in gynecology. Akush Ginecol Sofiia. 1995; 34: 14-16.
10. Mahony JB,Chernesky MA, Mores M. Detection of Chlamydia trachomatis antibody by EIA and IF in genital speciment for symptomatic and asymptomatic women and men. J Infect Dis 1986; 154: 141-8.
11. Witkin SS, Ledger WJ. Antibodies to Chlamydia trachomatis in sera of women with recurrent spontaneous abortion. Am J Obstet Gynecol 1992;
167: 135-139.
12. Gogate A, Deodhar L, Vaidya S. Detection of Chlamydia trachomatis Ab and toxoplasma gondii (IgG) and Mycoplasma hominis (IgG) in women with
bad obstetric history Ind J Med Res 1989; 100: 19- 22.
13. Maitrayee RM, Mathai M. Prevalence of genital Mycoplasma and Ureaplasma infections in pregnancy and their effect on pregnancy outcome Ind Med Res 1994; 100: 15-18.
14. Nancy EJ, Ruth B. Histology and Ureaplasma urealyticum culture in 63 case of first trimester abortion. Microbiol Infect Dis 1994; 6: 732-734.
15. Berg TG, Philpot KL, Welsh MS, Sanger WG, Ureaplasma. Mycoplasma infected amniotic fluid. J perinatol 1999; 19: 275-277.
16. Amortegul AJ, Meyer MP, Gnatuk CL.
Prevalence of Chlamydia trachomatis in women seeking abortion in Pittsbargh, Pennsylvania, United States of America. Genitourin Med 1986; 62:
88-92.
