Sero-detection of Chlamydia trachomatis Antibodies among Pregnant Women, Wad Medani Maternity Teaching Hospital, Gezira State, Sudan (2017) Mona Osman Ahmed Ali B.Sc. in Medical Microbiology, Faculty of Medicine and Health Sciences, Shendi University (2011) A Dissertation Submitted to University of Gezira in Partial Fulfillment for the Requirements for the Award of the Degree of Master of Laboratory Science in Medical Microbiology Department of Medical Microbiology Faculty of Medical Laboratory Sciences May, 2018 i Sero-detection of Chlamydia trachomatis Antibodies among Pregnant Women, Wad Medani Maternity Teaching Hospital, Gezira State, Sudan (2017) Mona Osman Ahmed Ali Supervision Committee: Name : Position Signature Dr: Nadir Musa Khalil Abuzeid Main supervisor …………. Dr: Adam Dawoud Abakar Co-supervisor …………. Date: …/…/2018 ii Sero-detection of Chlamydia trachomatis Antibodies among Pregnant Women, Wad Medani Maternity Teaching Hospital, Gezira State, Sudan (2017) Mona Osman Ahmed Ali Examination Committee: Name Position Signature Dr: Nadir Musa Khalil Abuzeid Chairperson …..……. Dr: Wad-Elbehar Hamad Elneel External examiner ...…….... Dr: Sanaa Elfatih Husain Internal examiner ………… Date: …/…/2018 iii Declaration I authorized that my dissertation” Sero-detection of Chlamydia trachomatis Antibodies among Pregnant Women, Wad Medani Maternity Teaching Hospital, Gezira State, Sudan (2017)" submitted by me under the supervision of Dr: Nadir Musa Khalil Abuzeid and Prof. Adam Dawoud Abakar for the partial fulfillment for the award of Master degree in Medical Laboratory Sciences in Medical Microbiology. University of Gezira Faculty of Medical Laboratory Sciences Department of Medical Microbiology; Wad- Medani, Sudan and this is original and it was not submitted in part or in full, in any printed or electronic means, and is not being considered elsewhere for publication or submission awarding degree elsewhere. Name and Signature of Candidate Name: Mona Osman Ahmed Ali Signature: ……………………….… Place: Wad Madani Date: …/…/2018 iv Dedication To my parents; thanks for supporting me; Thanks for giving me a chance to prove and improve myself through all my path of life. To my brothers, sisters, husband, and my daughters. To my friends and all people support me. And of course, all my colleagues in master program Batch (3). If I forgot anyone, please don’t ever doubt my dedication and love for ALL. v Acknowledgements I am grateful to all of helped me in completing this dissertation and provided me with information and special thanks and appreciation to my supervisors: Dr. Nadir Musa Khalil Abuzeid, Assistant Professor and consultant of medical microbiology University of Omdurman Islamic university, Faculty of Medical Laboratory Sciences, Department of Medical microbiology who followed and supported me in all Stages of this dissertation. I deeply thanks to my Co-supervisor Prof. Adam Dawoud Abakar Salim for hir guidance and great help. I do appreciate effort of all Staff working in Medical Microbiology and Post graduate studies departments. Especial thank to all staff of Wad Madani Obstetrics and Gynecology Hospital For collaboration during data collection. My particular thanks to my best friends Alaa Faroge , Awad Elymani, Ahmed omer and all staff of the faculty of medical laboratory sciences in Gezira University. Finally I am grateful to all who helped me and dedicated his time and effort to help me and did not mentioned hear above. vi Sero-detection of Chlamydia trachomatis Antibodies among Pregnant Women, Wad Medani Maternity Teaching Hospital, Gezira State, Sudan (2017) Mona Osman Ahmed Ali Abstract Chlamydia trachomatis is an obligate intracellular microorganism; it has ability to form intra cytoplasmic inclusion bodies within infected cells. In pregnant women, colonization of Chlamydia trachomatis in the genital tract during early gestation has been associated with spontaneous preterm birth. This case control hospital based study aims to detect Chlamydia trachomatis antibodies (IgG and IgM) among spontaneous recurrent miscarriage and non- miscarriage women at Wad Medani Maternity Hospital, Gezira State. A total of 90 women with and without history of miscarriage were enrolled in this study. 45(50%) were women with recurrent miscarriage, while the other 45(50 %) were women without history of miscarriage. Five ml of blood specimens were collected from each woman and dispensed into Ethylene Diamine Tetra Acetic acid and plain containers. Serum was collected by centrifugation at 4000 r/min for 10 minutes. Sera were tested for the presence of Chlamydia trachomatis IgG antibodies and Chlamydia trachomatis IgM antibodies using indirect Enzyme Linked Immunosorbent Assay (ELISA) method, Demographic and clinical data were collected by structured questionnaire after a verbal consent. 3/45 (6.7 %) women of study subjects were positive for Chlamydia trachomatis IgG antibodies and 3/45(6.7%) were positive for Chlamydia trachomatis IgM antibodies, while 2(2.2) women of control group were positive for Chlamydia trachomatis IgM antibodies. The study concluded that seroprevalence rate of Chlamydia trachomatis IgG and IgM antibodies are higher among women with recurrent miscarriage than those without history of recurrent miscarriage. The study recommended the screening for pregnant women for Chlamydia trachomatis. vii الكشف المصلي عن اﻻجسام المضادة للمتدثرة الحثرية بين النساء الحوامل, مستشفي ود مدني التعليمي للنساء والتوليد , وﻻية الجزيرة, السودان )2017م( منى عثمان احمد علي ملخص الدراسة المتدثرة الحثرية هي من الكائنات الحية الدقيقة التي تعيش داخل الخﻻيا. لديها القدرة على تشكيل جسيمات داخل الخﻻيا المصابة. في النساء الحوامل ارتبط وجود المتدثرة الحثرية في الجهاز التناسلي أثناء الحمل بالوﻻدة المبكرة التلقائية. تهدف دراسة الحالة والشاهد المستندة إلى المستشفي إلى الكشف عن المتدثرة الحثرية بين النساء الﻻتي أصبن باﻻجهاض والنساء الﻻتي لم يصبن باﻻجهاض التلقائي المتكرر المتردات علي مستشفي ود مدني للنساء والتوليد, وﻻية الجزيرة. شملت الدراسة 90امرأة , 45 )50 ٪( كن لديهم اجهاض ، 45 آخرين )50 ٪( من النساء ليس لديهم تاريخ اﻹجهاض. تم جمع خمسة مل من عينات الدم من كل امرأة وتم توزيعها في انابيب اختبار تحتوى علي مضاد التجلط ايثايلين دايمين تترا اثيدك اسيد واخرى ﻻ تحتوي علي مانع تجلط . تم تحضير المصل بواسطة الطرد المركزي عند 4000 لفة / دقيقة لمدة 10 دقائق. تم اختبار اﻷمصال لوجود اﻷجسام المضادة )ج( و اﻷجسام المضادة )م( للمتدثرة الحثرية عن طريق استخدم طريقة الفحص المناعي المرتبط باﻹنزيم غير المباشر ، وتم جمع البيانات الديموغرافية والسريرية من خﻻل استبيان بعد الحصول على الموافقة شفهيا من المستهدفين. من بين 45 امراة لديها اجهاض كانت 3 نساء )6.7%( نتائجهم ايجابية لـﻻجسام المضادة )ج( للمتدثرة الحثرية. و ثﻻث نساء )6.7٪( كانت نتائجهم ايجابية لﻷجسام المضاد )م( للمتدثرة الحثرية. بينما اثنان )4.4%( من النساء الﻻتي لم يصبن باﻻجهاض كانت نتائجهم ايجابية لﻷجسام المضادة ( م( للمتدثرة الحثرية .وخلصت الدراسة الي أن معدل انتشار اﻻجسام المضادة )ج , م( للمتدثرة الحثرية وسط النساء الﻻتي تعرضن لﻻجهاض اعلي من معدل اﻻنتشار وسط النساء الﻻتي ليس لديهن تاريخ إجهاض. اوصت الدراسة بأهمية اجراء فحص روتيني للكشف عن المتدثرة الحثرية لكل النساء الحوامل . viii TABLE OF CONTENTS Number CONTENTS Page Supervision committee i Examination committee ii Declaration iii Dedication iv Acknowledgement v English abstract vi Arabic abstract vi Table of contents Vii List of tables viii List of abbreviations ix CHAPTER ONE 1.1 Introduction 1 1.2 Justification 2 1.3 Objectives 2 1.3.1 General objective 2 1.3.2 Specific Objectives 2 CHAPTER TWO LITERATURE REVIEW 2.1 Definition of miscarriage 4 2.2 Classification of miscarriage 4 2.3 Causes of miscarriage 4 2.4 Chlamydia Trachomatis 5 2.4.1 Taxonomy 5 2.4.2 General properties 5 2.4.3 Morphology 6 2.4.4 Genomics 7 2.4.5 Life cycle 7 2.4.6 Antigenicity 8 2.4.7 Pathogenesis and Immunity 8 2.4.8 Clinical Syndromes 9 2.4.8.1 Lymphogranuloma Venereum 10 2.4.8.2 Ocular genital Infections 10 2.4.8.3 Urogenital infections 10 2.4.8.4 Trachoma 11 ix 2.4.8.5 Neonatal pneumonia 11 2.4.8.6 Fitz-Hugh-Curtis 12 2.4.9 laboratory Diagnosis 12 2.4.9.1 Specimens 12 2.4.9.2 diagnostic methods 12 2.4.9.2.1 Direct cytological examination 12 2.4.9.2.2 Isolation in cell culture 13 2.4.9.2.3 Antigen Detection 13 2.4.9.2.4 Nucleic Acid Hybridization 13 2.4.9.2.5 Nucleic Acid Amplification Tests 14 2.4.9.2.6 Serodiagnosis 14 2.4.9.3 Antimicrobial susceptibility testing 15 2.4.10 Treatment 15 2.4.11 prevention and control 15 2.5 The Relationship between Chlamydia trachomatis Genital Infection 16 and Spontaneous Abortion 2.6 Previous studies 16 CHAPTER THREE (MATERIALS AND METHODS) 3.1 Study design 18 3.2 Study area 18 3.3 Study population 18 3.4 Sample Size 18 3/5 Study Criteria 18 3.5.1 Inclusion criteria 18 3.5.2 Exclusion criteria 19 3.6 Data collection 19 3.7 Data analysis 19 3.8 Ethical consideration 19 3.9 Laboratory method 19 3.9.1 Collection of blood specimens 19 3.9.2 Sample processing 19 3.9.3 CBCs processing 19 3.9.4 Enzyme linked immunosorbent assay (ELISA) processing 20 x 3.9.4.1 Principle of Enzyme linked immunosorbent assay (ELISA) for 20 detection of C. trachomatis IgG and IgM antibodies 3.9.4.2 Procedure 20 3.9.4.3 Quality control and calculation of results 21 CHAPTER FOUR 4.1 Results 22 4.2 Discussion 32 CHAPTER FIVE 5.1 Conclusion 35 5.2 Recommendations 35 References 36 Appendix 42 xi LIST OF TABLE Table Title Page 4.1 Socio-demogric and clinical characteristic of study population 23 4.2 Serodetection of anti C. trachomatis IgG and IgM according to Miscarriage 24 4.3 Serodetection of anti C. trachomatis IgG and IgM according to rate of miscarriage 24 4.4 Serodetection of anti C. trachomatis IgG and IgM according to trimester: 24 4.5 Serodetection of anti C. trachomatis IgG and IgM according to family history 25 4.6 Serodetection of anti C.