RESUMO
Human CMV is the most causative agent of a very common viral infection contacted by most adults that have no noticeable or with only mild uncharacteristic symptoms. However when a pregnant women is infected with CMV as a primary infection, there is a risk for transmission of virus to the fetus as well as reactivation of virus in rare case. HCMV antibodies were already described in spontaneous abortion and fetal abnormalities cases. Also antibodies against HCMV in fetal abnormalities as well as abortion had been reported by several studies in different part of the world. Due to lack of published data about CMV epidemiology in Ilam, the aim of current study was to determine the seroprevalence of specific viral IgM and IgG in spontaneous abortion cases as well as the age and socioeconomic status in the studied population in Ilam. Sera sample from 42 patients in abortion process as well as 30 healthy pregnant and 30 healthy women as negative control were collected and quantitative serological test to assess IgM/IgG against HCMV was performed using a commercial ELISA assay. SPSS software was used to analysis the results and demographic information. Among 42 patients in abortion process, IgG was found in 6 [14.28%] patients and IgM in 12 [28.58%] cases. Based on demographical information, it was showed that IgG seropositivity correlate with the increase of age, but there is no correlation between IgM and age of patients. The results showed that there is a high seroPrevalence of HCMV IgM than IgG among pregnant women in the process of abortion in Ilam; correlation between Age and IgG anti body seroPrevalence was same as other reported. Based on the current studies, it seems that more sensitive and specific method such as NAT method is needed for determination of CMV and abortion procces
RESUMO
Polymerase chain reaction [PCR] is one of the most important progress in the field of molecular biology and diagnosis. Despite simplicity in concept, the reaction needs complex interaction between target sequence, primers, dNTPs and DNA polymerase for a successful amplification and diagnosis. For the detection of RNA viruses highly sensitive and specific technique is required. Hence amplification based on Nested PCR would improve sensitivity, and also use of one step reaction would decrease probable contaminations as reported previously in several studies. The aim of the current study was to develop a new, rapid and sensitive one step-one tube Nested PCR in a closed system, by using two novel coherent primers. In this study, a novel and special primer development method was used for one step-one tube reaction. After development and optimization, the assay was evaluated with known positive and negative controls. The developed assay was performed on 50 HCV positive samples and 10 negative controls and 5 samples from each HIV, HBV, TTV [Torque Teno Virus] and GBV-C [Hepatitis G Virus: HGV]. Based on the obtained results, sensitivity and specifity was calculated. 48 out of 50 HCV positive samples showed expected band while none of the negative controls gave any band. Based on the specific primer design system which has been used in the current study; the inner primer was synthesized as complementary of routine PCR primers and was bound to the outer primers. Therefore, there is no probability for false priming in the both rounds of the reaction, hence there would be no nonspecific amplifications. Other advantages of this assay system were prevention of contamination which was due to one step-one tube reaction, decrease in duration and the cost of the reaction