Brief Communication Isolation of Mycoplasma genitalium from endocervical swabs of infertile women Dunia A. Alfarraj, PhD, Ali M. Somily, MD, FRCPC.
ABSTRACT Objectives: To determine the occurrence of Mycoplasma genitalium (M. genitalium) infection in infertile women attending infertility clinic and its association to infertility. Methods: Endocervical specimens were collected from women presenting with primary and secondary infertility and from fertile women as control group. Real-time polymerase chain reaction (PCR) assay was performed to detect the presence of M. genitalium from these endocervical specimens. Results: Mycoplasma genitalium was detected in 3% of infertile women. There was no statistically significant difference between infertility and control group as to signs and symptoms except for signs of cervicitis were presented only among infertile group. We found no significant differences among fertile and infertile women for M. genitalium infection. Conclusion: An association between M. genitalium infection and infertility may exist. Screening of women for M. genitalium infection is recommended as part of investigations for infertility. Saudi Med J 2017; Vol. 38 (5): 549-552 doi: 10.15537/smj.2017.5.18820
I
nfertility is a worldwide health problem with a current global infertility of 15%.1 In approximately 10% to 15% of couples, no direct cause of their infertility can be found, and 20% - 40% of couples may have multiple factors for infertility.2 Approximately 35% of infertile women are afflicted with post-inflammatory changes of the oviduct or surrounding peritoneum that interfere with tubal-ovarian function mostly as a result from infection and are likely to develop an ectopic (tubal) pregnancy.3 Mycoplasma genitalium (M. genitalium) is a small parasitic bacterium that lives on various cells of the body including the genital and respiratory tracts, fallopian tube cells, and spermatozoa.4 The human urogenital tract is the preferred site of colonization, which strongly indicates
OPEN ACCESS
that M. genitalium is sexually transmitted.5 Mycoplasma genitalium was detected in mid-trimester of women delivering preterm,6 and among women with tubal factor infertility.7 Immunological assays cannot reliably detect the presence of M. genitalium.8 Currently, several tests are being used to detect Mycoplasma infections including nucleic acid amplification testing (NAATs), and a quantitative real-time Light Cycler polymerase chain reaction (PCR) assay which targets the gene gap encoding glyceraldehyde-3-phosphate dehydrogenase.9 In Saudi Arabia, discussing issues about sexually transmitted infections (STIs) is considered taboo where ethics and social factors give rise to many obstacles, thus there is a possibility that STIs might pose a significant public health threat. Data of M. genitalium infections are limited in Saudi Arabia. This study was conducted to determine the occurrence of M. genitalium among infertile women, compare it with a control of fertile women, evaluate its association with the type of infertility, infertility factors and their occurrence, and established a convenient method for rapid detection and identification of M. genitalium infection among infertile women. Methods. Between October 2012 and July 2013, this prospective case control study was conducted in King Khalid University Hospital (KKUH) and King Abdulaziz University Hospital (KAUH), both in Riyadh, Saudi Arabia. Ethical consent was obtained from the ethical committee board of KKUH. One hundred infertile married women with primary and secondary infertility aged between 19 and 46 years who attended the outpatient infertility clinic at KKUH for infertility examination and fulfilled the inclusion criteria during the period of study and agreed to participate were enrolled and formed the case group. Another group of 100 women who attended the clinics for gynecologic purposes, either for routine or annual check-up comprised the control group. Relevant medical records were reviewed for any possible present and past medical or surgical diseases. Pregnant women and women who had taken antibiotics in the previous 30 days were excluded from this study. Participants from both groups completed a questionnaire form on clinico-demographic information. Gynecological data including symptomatologies of gynecologic infection were obtained and recorded. Endocervical swabs by a speculum examination were
www.smj.org.sa
Saudi Med J 2017; Vol. 38 (5)
549
M. genitalium and infertility in women ... Alfarraj & Somily
collected from all participants. Swabs were stored at 4°C until transported to the laboratory, and stored at 20°C until processed. Deoxyribonucleic acid were extracted using the MagNA Pure Compact Nucleic Acid Isolation Kit (Roche Diagnostics and MagNA Pure Compact system, Mannheim, Germany), and was processed according to manufacturers protocol. Deoxyribonucleic acid amplification and detection protocol was carried out using the Light Cycler Fast Start DNA Master HybProbe according to the manufacturers protocol. PCR assays were carried out using the LightCycler PCR program, and melting curves were obtained according to the LightCycler instrument operator’s manual (TIB Molbiol, Germany). Statistical analysis were performed using Statistical Package for Social Sciences (SPSS) version 19.0 (IBM Corp., Armonk, NY, USA). Results are presented as frequencies and percentages for categorical variables, and mean and standard deviation for numerical variables. We used chi-square test to compare between infertile group and the non-infertile group, and to compare between primary infertility and secondary infertility with respect to all nominal variables. Correlations were carried out using the Pearson correlation. P values of
ABSTRACT Objectives: To determine the occurrence of Mycoplasma genitalium (M. genitalium) infection in infertile women attending infertility clinic and its association to infertility. Methods: Endocervical specimens were collected from women presenting with primary and secondary infertility and from fertile women as control group. Real-time polymerase chain reaction (PCR) assay was performed to detect the presence of M. genitalium from these endocervical specimens. Results: Mycoplasma genitalium was detected in 3% of infertile women. There was no statistically significant difference between infertility and control group as to signs and symptoms except for signs of cervicitis were presented only among infertile group. We found no significant differences among fertile and infertile women for M. genitalium infection. Conclusion: An association between M. genitalium infection and infertility may exist. Screening of women for M. genitalium infection is recommended as part of investigations for infertility. Saudi Med J 2017; Vol. 38 (5): 549-552 doi: 10.15537/smj.2017.5.18820
I
nfertility is a worldwide health problem with a current global infertility of 15%.1 In approximately 10% to 15% of couples, no direct cause of their infertility can be found, and 20% - 40% of couples may have multiple factors for infertility.2 Approximately 35% of infertile women are afflicted with post-inflammatory changes of the oviduct or surrounding peritoneum that interfere with tubal-ovarian function mostly as a result from infection and are likely to develop an ectopic (tubal) pregnancy.3 Mycoplasma genitalium (M. genitalium) is a small parasitic bacterium that lives on various cells of the body including the genital and respiratory tracts, fallopian tube cells, and spermatozoa.4 The human urogenital tract is the preferred site of colonization, which strongly indicates
OPEN ACCESS
that M. genitalium is sexually transmitted.5 Mycoplasma genitalium was detected in mid-trimester of women delivering preterm,6 and among women with tubal factor infertility.7 Immunological assays cannot reliably detect the presence of M. genitalium.8 Currently, several tests are being used to detect Mycoplasma infections including nucleic acid amplification testing (NAATs), and a quantitative real-time Light Cycler polymerase chain reaction (PCR) assay which targets the gene gap encoding glyceraldehyde-3-phosphate dehydrogenase.9 In Saudi Arabia, discussing issues about sexually transmitted infections (STIs) is considered taboo where ethics and social factors give rise to many obstacles, thus there is a possibility that STIs might pose a significant public health threat. Data of M. genitalium infections are limited in Saudi Arabia. This study was conducted to determine the occurrence of M. genitalium among infertile women, compare it with a control of fertile women, evaluate its association with the type of infertility, infertility factors and their occurrence, and established a convenient method for rapid detection and identification of M. genitalium infection among infertile women. Methods. Between October 2012 and July 2013, this prospective case control study was conducted in King Khalid University Hospital (KKUH) and King Abdulaziz University Hospital (KAUH), both in Riyadh, Saudi Arabia. Ethical consent was obtained from the ethical committee board of KKUH. One hundred infertile married women with primary and secondary infertility aged between 19 and 46 years who attended the outpatient infertility clinic at KKUH for infertility examination and fulfilled the inclusion criteria during the period of study and agreed to participate were enrolled and formed the case group. Another group of 100 women who attended the clinics for gynecologic purposes, either for routine or annual check-up comprised the control group. Relevant medical records were reviewed for any possible present and past medical or surgical diseases. Pregnant women and women who had taken antibiotics in the previous 30 days were excluded from this study. Participants from both groups completed a questionnaire form on clinico-demographic information. Gynecological data including symptomatologies of gynecologic infection were obtained and recorded. Endocervical swabs by a speculum examination were
www.smj.org.sa
Saudi Med J 2017; Vol. 38 (5)
549
M. genitalium and infertility in women ... Alfarraj & Somily
collected from all participants. Swabs were stored at 4°C until transported to the laboratory, and stored at 20°C until processed. Deoxyribonucleic acid were extracted using the MagNA Pure Compact Nucleic Acid Isolation Kit (Roche Diagnostics and MagNA Pure Compact system, Mannheim, Germany), and was processed according to manufacturers protocol. Deoxyribonucleic acid amplification and detection protocol was carried out using the Light Cycler Fast Start DNA Master HybProbe according to the manufacturers protocol. PCR assays were carried out using the LightCycler PCR program, and melting curves were obtained according to the LightCycler instrument operator’s manual (TIB Molbiol, Germany). Statistical analysis were performed using Statistical Package for Social Sciences (SPSS) version 19.0 (IBM Corp., Armonk, NY, USA). Results are presented as frequencies and percentages for categorical variables, and mean and standard deviation for numerical variables. We used chi-square test to compare between infertile group and the non-infertile group, and to compare between primary infertility and secondary infertility with respect to all nominal variables. Correlations were carried out using the Pearson correlation. P values of
