Molecular detection of intrauterine microbial colonization in women with endometriosis

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Reviewed Marked as Reviewed by Atrayees on 2023-8-7
study design
Citation
PMID PubMed identifier for scientific articles.
DOI Digital object identifier for electronic documents.
URI
Authors
Khan KN, Fujishita A, Masumoto H, Muto H, Kitajima M, Masuzaki H, Kitawaki J
Journal
European journal of obstetrics, gynecology, and reproductive biology
Year
2016
Keywords:
16S rDNA metagenome assay, Cystic fluid, Endometriosis, Infection
OBJECTIVE: Increased intrauterine microbial colonization by bacteria culture method and occurrence of endometritis have been reported in women with endometriosis. Here we investigated microbial colonization in intrauterine environment and cystic fluid of women with and without endometriosis by molecular approach. STUDY DESIGN: This is a case-controlled biological study with a total of 32 women each with and without endometriosis. Among them, 16 each in these two groups of women received treatment with gonadotropin-releasing hormone agonist (GnRHa). Pattern of microbial colonization in endometrial swabs and endometrioma/non-endometrioma cystic fluid was examined using broad-range polymerase-chain reaction (PCR) amplification of bacteria targeting 16S rRNA gene (rDNA). After quantification of index PCR product, 16S rDNA metagenome sequence analysis was done by Illumina Miseq system. RESULTS: A wide proportion (0.01-97.8%) of multiple bacteria was detected in both endometrial swabs and cystic fluid collected from women with and without endometriosis. 16S metagenome assay indicated that proportion of Lactobacillacae was significantly decreased (p<0.01) and of Streptococcaceae, Staphylococaceae, Enterobacteriaceae was significantly increased (p<0.05 for each) in GnRHa-treated women with endometriosis than in GnRHa-untreated women. While bacteria culture method failed to detect a single colony, 16S metagenome assay could detect significantly higher percentage of Streptococcaceae (p<0.01) and Staphylococaceae (p<0.05) in the cystic fluid derived from women with ovarian endometrioma comparing to that in cystic fluid collected from non-endometrioma cysts. CONCLUSION: These findings indicate the occurrence of sub-clinical infection in intrauterine environment and in the cystic fluid of ovarian endometrioma. Additional side effect of GnRHa treatment in promoting silent intrauterine and/or ovarian infection should be considered.

Experiment 1


Reviewed Marked as Reviewed by Atrayees on 2023-8-7

Curated date: 2021/08/08

Curator: Samara.Khan

Revision editor(s): WikiWorks, Samara.Khan, Atrayees, Peace Sandy

Subjects

Location of subjects
Japan
Host species Species from which microbiome was sampled. Contact us to have more species added.
Homo sapiens
Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology
Endometrium Tunica mucosa (endometrium),Tunica mucosa uteri,Uterine endometrium,Uterine mucosa,Uterine mucous membrane,Endometrium,endometrium
Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Endometriosis endometriosis,Endometriosis (clinical),endometriosis (disease),Endometriosis (disorder),Endometriosis (morphologic abnormality),ENDOMETRIOSIS NEC,Endometriosis NOS,Endometriosis NOS (disorder),Endometriosis of other specified sites,Endometriosis, site unspecified,Endometriosis
Group 0 name Corresponds to the control (unexposed) group for case-control studies
Untreated control women
Group 1 name Corresponds to the case (exposed) group for case-control studies
Endometriosis patients
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
Patients were diagnosed with endometriosis via laparoscopic surgery and stages were categorized according to the revised American Society for Reproductive Medicine scoring system (r-ASRM). 16 of the control patients and 16 of the endometriosis patients received GnRHa (gonadotropin releasing hormone agonist) treatment for 4-6 months.
Group 0 sample size Number of subjects in the control (unexposed) group
32
Group 1 sample size Number of subjects in the case (exposed) group
32
Antibiotics exclusion Number of days without antibiotics usage (if applicable) and other antibiotics-related criteria used to exclude participants (if any)
None

Lab analysis

Sequencing type
16S
16S variable region One or more hypervariable region(s) of the bacterial 16S gene
Not specified
Sequencing platform Manufacturer and experimental platform used for quantifying microbial abundance
Illumina

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).
relative abundances
Statistical test
Mann-Whitney (Wilcoxon)
Significance threshold p-value or FDR threshold used for differential abundance testing (if any)
0.05
MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?
No


Signature 1

Reviewed Marked as Reviewed by Atrayees on 2023-8-7

Curated date: 2021/08/08

Curator: Samara.Khan

Revision editor(s): Samara.Khan

Source: Figure 3

Description: Regardless of GnRHa treatment, patients with endometriosis had higher levels of streptococcaceae and moraxellaceae than those without endometriosis.

Abundance in Group 1: increased abundance in Endometriosis patients

NCBI Quality ControlLinks
Streptococcaceae
Moraxellaceae

Revision editor(s): Samara.Khan

Experiment 2


Reviewed Marked as Reviewed by Atrayees on 2023-8-7

Curated date: 2021/08/08

Curator: Samara.Khan

Revision editor(s): Samara.Khan, WikiWorks

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies
Endometriosis patients not treated with GnRHa
Group 1 name Corresponds to the case (exposed) group for case-control studies
Endometriosis patients treated with GnRHa
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
Patients were diagnosed with endometriosis via laparoscopic surgery and stages were categorized according to the revised American Society for Reproductive Medicine scoring system (r-ASRM). 16 of the 32 endometriosis patients received GnRHa (gonadotropin releasing hormone agonist) treatment for 4-6 months.
Group 0 sample size Number of subjects in the control (unexposed) group
16
Group 1 sample size Number of subjects in the case (exposed) group
16

Lab analysis

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).
Not specified


Signature 1

Reviewed Marked as Reviewed by Atrayees on 2023-8-7

Curated date: 2021/08/08

Curator: Samara.Khan

Revision editor(s): Samara.Khan

Source: Figure 4

Description: Streptococcaceae, Staphylococaceae, Enterobacteriaceae were significantly increased (p<0.05 for each) in GnRHa-treated women with endometriosis than in GnRHa-untreated women.

Abundance in Group 1: increased abundance in Endometriosis patients treated with GnRHa

NCBI Quality ControlLinks
Enterobacteriaceae
Streptococcaceae
Staphylococcaceae

Revision editor(s): Samara.Khan

Signature 2

Reviewed Marked as Reviewed by Atrayees on 2023-8-7

Curated date: 2021/08/08

Curator: Samara.Khan

Revision editor(s): Samara.Khan

Source: Figure 4

Description: Lactobacillaceae was significantly decreased (p<0.01) in GnRHa-treated women with endometriosis compared to GnRHa-untreated women with endometriosis

Abundance in Group 1: decreased abundance in Endometriosis patients treated with GnRHa

NCBI Quality ControlLinks
Lactobacillaceae

Revision editor(s): Samara.Khan

Experiment 3


Reviewed Marked as Reviewed by Atrayees on 2023-8-7

Curated date: 2021/08/08

Curator: Samara.Khan

Revision editor(s): Samara.Khan, WikiWorks

Differences from previous experiment shown

Subjects

Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology
Ovary Animal ovary,Female gonad,Female organism genitalia gonad,Female organism genitalia gonada,Female organism reproductive system gonad,Female organism reproductive system gonada,Female reproductive system gonad,Female reproductive system gonada,Genitalia of female organism gonad,Genitalia of female organism gonada,Gonad of female organism genitalia,Gonad of female organism reproductive system,Gonad of female reproductive system,Gonad of genitalia of female organism,Gonad of reproductive system of female organism,Gonada of female organism genitalia,Gonada of female organism reproductive system,Gonada of female reproductive system,Gonada of genitalia of female organism,Gonada of reproductive system of female organism,Ovaries,Ovarium,Ovum-producing ovary,Reproductive system of female organism gonad,Reproductive system of female organism gonada,Ovary,ovary
Group 0 name Corresponds to the control (unexposed) group for case-control studies
Patients with non-endometrioma cysts
Group 1 name Corresponds to the case (exposed) group for case-control studies
Patients with ovarian endometrioma cysts
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
Patients with ovarian endometrioma had endometrial cysts in their ovaries. Researchers took samples of the fluid inside these cysts. Ovarian endometrioma is one of the many ways endometriosis manifests in the female reproductive system.
Group 0 sample size Number of subjects in the control (unexposed) group
8
Group 1 sample size Number of subjects in the case (exposed) group
8

Lab analysis

Statistical Analysis

Signature 1

Reviewed Marked as Reviewed by Atrayees on 2023-8-7

Curated date: 2021/08/08

Curator: Samara.Khan

Revision editor(s): Samara.Khan

Source: Figure 5

Description: There was a significantly higher percentage of Streptococcaceae (p<0.01) and Staphylococaceae (p<0.05) in the cystic fluid derived from women with ovarian endometrioma comparing to that in cystic fluid collected from non-endometrioma cysts.

Abundance in Group 1: increased abundance in Patients with ovarian endometrioma cysts

NCBI Quality ControlLinks
Streptococcaceae
Staphylococcaceae

Revision editor(s): Samara.Khan

Signature 2

Reviewed Marked as Reviewed by Atrayees on 2023-8-7

Curated date: 2023/08/07

Curator: Atrayees

Revision editor(s): Atrayees

Source: Figure 5

Description: There was a significantly higher percentage of Streptococcaceae (p<0.01) and Staphylococaceae (p<0.05) in the cystic fluid derived from women with ovarian endometrioma comparing to that in cystic fluid collected from non-endometrioma cysts.

Abundance in Group 1: decreased abundance in Patients with ovarian endometrioma cysts

NCBI Quality ControlLinks
Lactobacillaceae

Revision editor(s): Atrayees