Molecular Detection of Cytomegalovirus (CMV) Isolated from Repeated Miscarriage Women and its Relation to TLR2 and ILT2 Receptors

number: 
3528
إنجليزية
Degree: 
Author: 
Sara Ahmed Jihad
Supervisor: 
Dr. Rehab Subhi Ramadhan(Assist. prof.)
year: 
2015

      This study was designed to determine a molecular status of Toll like Receptor 2 in repeated miscarriage women with cytomegalovirus. One hundred blood samples were collected from repeated miscarriage women with cytomegalovirus from infertility clinic of Kamal Al –Sammaraee hospital and (50) samples from normal subjects served as control for comparison and divided into two age groups:( 20-30) and (31-40) years old. Women distributed as (60) samples of infertile and (40) samples as miscarriage women.The first part of this study: anti- HCMV antibodies IgG and IgM was estimated by Enzyme Linked Immune Sorbent Assay (ELISA). Results showed that the miscarriage women showed the highest percentage of seropositive to CMV which were 40%of IgG and 25% of IgM compared with infertile women that showed 20% of IgG and 15% of IgM with a significant difference P ≤ 0.05. The Seropositive of anti- HCMV IgG was higher in younger women (20-30) years old, while the age group (31-40) years showed higher level of IgM. The second part of this study included molecular analysis using polymerase chain reaction technique (PCR) for amplification two viral genes which are glycoprotein gB gene, that showed molecular size 72bp and immediate early IE2 gene that showed band with molecular size 92bp. Real time PCR used for diagnosing CMV infection by gB-primer with probe FAM and TAMRA, the result showed amplification from cycle six. The amplification for TLR2 primer region showed band with molecular size 285bp. After successful amplification of TLR2 gene, good quality products were selected to be sequenced. The result of TLR2 sequencing showed only replacement of C instead of G in wild type at position (242).PCR amplified for ILT2 primer region showed band with molecular size of419bp. Among ten Iraqi patients, six only give positive result whencompared with healthy control; the type of mutation is deletion andsubstitution. The percentage of mutation types were deletion mutation13.3% and 86.7% for substitution mutation. The effect of mutation wasmissense mutation 72.7%, Silent mutations 15.1%, and deletion mutations12.2%.