{"id":13545,"date":"2017-03-25T11:06:27","date_gmt":"2017-03-25T11:06:27","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=13545"},"modified":"2020-04-23T09:49:12","modified_gmt":"2020-04-23T09:49:12","slug":"detection-of-cytomegalovirus-and-epstein-barr-virus-in-placental-tissues-of-aborted-women","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol10no1\/detection-of-cytomegalovirus-and-epstein-barr-virus-in-placental-tissues-of-aborted-women\/","title":{"rendered":"Detection of Cytomegalovirus and Epstein Barr Virus in Placental Tissues of Aborted Women"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

The abortions reasons in several circumstances yet are mysterious, nevertheless the bacterial toxicities signify a main reason in abortion, where germs seems to be the utmost elaborate pathogens (Khameneh et.al<\/em>., 2014) and (Oliver and Overton ,2014).<\/p>\n

Between numerous germs, Humanoid Herpes germ toxicities of placenta which could be hurtful in pregnant foremost to complaints in the growth of fatality, early delivery, failure, or main irregularities, besides it could yield to a prolonged or frequent parental infection. In specific, CMV through pregnancy could spread the placenta\u00a0\u00a0 by viremia, subsequent to main and recurring toxicity, or by rising way as of the cervix, regularly subsequent recurrence. The germ with a smallest mid herpes viruses, Epstein-Barr virus which usually related with irregular abortions (Avgil and Ornoy, 2006) and (Di Stefano et.al<\/em>., 2008).<\/p>\n

The goal of the current research is to investigate the differences in the presence of two herpetic virus-related toxicities inside placental soft tissue as of caseswhose spontaneously abortion through determination of infections with CMV, and EBV involving the placenta as a possible causes for subsequent abortion by using the following techniques: Immunohistochemistry and In situ hybridization; as well as comparing the two techniques for detection of CMV, and EBV.<\/p>\n

Method<\/strong><\/p>\n

This retrospective study made the use of paraffin contain placental soft tissue that composed from histo-pathological annals of Training Labs form the hospital of AL-Yarmouk in Baghdad \/ Iraq and belongs to forty (40) feminine cases with miscarriages as patient\u2019s collection; the range of the ages were (19-43 years), and 40 placental soft tissue of normal delivery as a control group. Depiction Rabbit and Mouse with certain HRP \\DAB inspection IHC Kit ab80436 (2013) Abcam was used for inspection of CMV \u2013protein and Epstein Barr – viral capsid antigen (EBV \u2013VCA) specific\u00a0 primary antibodies . CISH Implementation AP-NBT kit for detection of EBV (EBER \u2013 RNA) and CMV (DNA) for chromogenic in situ hybridization (CISH) using biotin \u2013 labeled Zyto Fast CISH probe was purchased from ZytoFast\/Germany Cat. Numbers (T-1070-40 – 2011). Numerical investigation: <\/strong>Investigation of statistics was achieved by means of the obtainable numerical set of (SPSS-22).<\/p>\n

Results<\/strong><\/p>\n

For IHC technique, the expression of EBV \u2013VCA IHC signs were identified as a brown color staining localized at nuclear (Figure1, A). The placental soft tissue tasters of aborted women presented 22.5% (9 cases from 40), whereas not any of well control placental soft tissue displayed EBV – VCA antigen expression. The maximum ratio of EBV-IHC shows a modest sign 66. 7 % (6 cases from 9) \u00a0as depicted in Table 1.<\/p>\n\n\n\n
\u00a0\"Figure<\/td>\nFigure 1: Microphotograph staining of <\/strong>trophoblastic <\/strong>placental <\/strong>tissues<\/strong> from miscarriage patients (red arrow): <\/strong>A-<\/strong> EBV<\/strong> IHC staining<\/strong> in <\/strong>cell nucleus intensity. <\/strong>B- <\/strong>IHC staining for <\/strong>CMV\u00a0\u00a0 <\/strong>at cell nucleus. C- Negative IHC staining for <\/strong>EBV. D-<\/strong> EBV-ERER\u00a0 CISH positive signals in the cell nucleus of <\/strong>\u00a0trophoblastic <\/strong>placental\u00a0 <\/strong>tissues <\/strong>\u00a0showed score 2 and moderate\u00a0 intensity. E-CMV -DNA staining CISH of <\/strong>trophoblastic <\/strong>placental <\/strong>tissues<\/strong> in the cell nucleus score 1 and weak intensity. F- Negative CISH staining for <\/strong>EBV<\/strong>.<\/strong><\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n


\nTable 1: Immuno-histochemical scoring & signal intensity data of EBV \u2013VCA
\n<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Groups<\/strong><\/p>\n

\u00a0<\/strong><\/p>\n<\/td>\n

Miscarriage<\/strong><\/p>\n

Set <\/strong><\/td>\n

Control<\/strong><\/p>\n

Set<\/strong><\/td>\n

P value<\/strong><\/td>\n<\/tr>\n
Number<\/strong><\/td>\n%<\/strong><\/td>\nNumber<\/strong><\/td>\n%<\/strong><\/td>\n<\/tr>\n
EBV IHC Positively Score<\/strong><\/td>\nNegatively<\/td>\n31<\/td>\n77.50<\/td>\n40.00<\/td>\n100.00<\/td>\n<\/td>\n<\/tr>\n
Positively<\/td>\n9<\/td>\n22.50<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
<\/td>\n(Score I)<\/td>\n7<\/td>\n77.80<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Score II)<\/td>\n2<\/td>\n22.20<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Score III)<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
EBV IHC Positively\u00a0 Intensity<\/strong><\/p>\n

 <\/td>\n

(Weakly \/ I)<\/td>\n3<\/td>\n33.30<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Moderately \/ II)<\/td>\n6<\/td>\n66.70<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Strongly \/ III)<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

The general appearance of CMV protein at nuclear localization (Figure1, B) was distinguished in 37.5% of the placental soft tissue of failure group and in 5% of healthy placental soft tissue. A great percent (around 60%) among placental soft tissue in the failure group had feeble score (score I). The uppermost positive\u00a0 CMV-IHC reactions in those with miscarriage group were showed strong\u00a0 signal intensity 66.7%\u00a0 .Statistically, noteworthy changes (p <0.05) was distinguished associating with the infection degree\u00a0 among miscarriage and control group (Table 2).<\/p>\n

Table 2<\/strong>: CMV-protein of IHC in placental soft tissue<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Groups<\/strong><\/p>\n

\u00a0<\/strong><\/p>\n<\/td>\n

Miscarriage<\/strong><\/p>\n

Set<\/strong><\/td>\n

Control<\/strong><\/p>\n

Group<\/strong><\/td>\n

P value<\/strong><\/td>\n<\/tr>\n
No.<\/strong><\/td>\n%<\/strong><\/td>\nNumber<\/strong><\/td>\n%<\/strong><\/td>\n\u00a0<\/strong><\/td>\n<\/tr>\n
CMV IHC Positively Score<\/strong><\/td>\nNegatively<\/td>\n25.00<\/td>\n62.50<\/td>\n38.00<\/td>\n95.00<\/td>\n1x 10-4<\/sup>*<\/td>\n<\/tr>\n
Positively<\/td>\n15.00<\/td>\n37.50<\/td>\n2.00<\/td>\n5.00<\/td>\n<\/td>\n<\/tr>\n
<\/td>\n(Score I)<\/td>\n9.00<\/td>\n60.00<\/td>\n2.00<\/td>\n100<\/td>\n<\/td>\n<\/tr>\n
(Score II)<\/td>\n1.00<\/td>\n6.70<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Score III)<\/td>\n5.00<\/td>\n33.30<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
CMVIHC Positively Intensity<\/strong><\/td>\n(Weakly \/ I)<\/td>\n1.00<\/td>\n6.70<\/td>\n2.00<\/td>\n100.00<\/td>\n<\/td>\n<\/tr>\n
(Moderately \/ II)<\/td>\n4.00<\/td>\n26.70<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Strongly \/ III)<\/td>\n10.00<\/td>\n66.70<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

*Very clear differences among the ratios by (Pearson Chi-square test at 0.05 level)<\/p>\n

The chromo-genic are in situ hybridization (CISH) system, the whole percent of positive CISH of EBERS between placental soft tissue in the miscarriage group was 22.5%, while none of the 40 control placental tissues have revealed positive CISH- signals for this EBV marker (figure 1, D). The scoring of EBER RNA- CISH reactions\u00a0 was in its highest percentage \u00a066.7% \u00a0of all positive cases\u00a0 in the low score (score I), while the their signal\u00a0 intensity of the\u00a0 color development where the percentage of\u00a0 EBER- RNA \u2013CISH reactions that showed weak, moderate, and strong\u00a0 intensities were 4(44.5%), 3(33.3%), and 2(22.2%) respectively (Table 3).<\/p>\n

Table 3: Distribution of scoring and intensity of CISH actions for EBER RNA- in placental tissues of miscarriage and control group<\/strong>.<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Groups<\/strong><\/p>\n

\u00a0<\/strong><\/p>\n<\/td>\n

Miscarriage<\/strong><\/p>\n

\u00a0Group<\/strong><\/td>\n

Control <\/strong><\/p>\n

Group<\/strong><\/td>\n

P value<\/strong><\/td>\n<\/tr>\n
No<\/strong><\/td>\n%<\/strong><\/td>\nNo<\/strong><\/td>\n%<\/strong><\/td>\n<\/tr>\n
\u00a0 EBV CISH Positively Score<\/strong><\/td>\nNegatively<\/td>\n31.00<\/td>\n77.50<\/td>\n40.0<\/td>\n100.00<\/td>\n<\/td>\n<\/tr>\n
Positively<\/td>\n9.00<\/td>\n22.50<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
<\/td>\n(Score I)<\/td>\n6<\/td>\n66.70<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Score II)<\/td>\n3<\/td>\n33.30<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Score III)<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
EBV CISH Positively Intensity<\/strong><\/p>\n

 <\/td>\n

\u00a0(Weakly \/ I)<\/td>\n\u00a0\u00a0 4<\/td>\n\u00a044.4<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Moderately \/ II)<\/td>\n\u00a0\u00a0 3<\/td>\n\u00a033.3<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
(Strong \/ III)<\/td>\n\u00a0\u00a0 2<\/td>\n\u00a022.2<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

CMV DNA was noticed in (30%) from the studied placental soft tissue in the miscarriage set, whereas placental tissues of healthy delivered women have revealed in (15%). Regarding the signal scoring, the most frequently detected CMV \u2013 positive\u00a0 placental tissue in the\u00a0 miscarriage group were observed to have the lower score (score I),<\/em> (50%), 58.3 % revealed\u00a0 weak\u00a0 sign strength ,numerically, no difference observed\u00a0\u00a0\u00a0 (p >0.05) as depicted in (Table 4).<\/p>\n

Table 4<\/strong>: Distribution of HCMV DNA CISH result of the examined placental tissues of miscarriage and their counterpart control tissues<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 CMV CISH signal Score &<\/strong>\u00a0Signal intensity<\/strong><\/td>\nMiscarriage Group<\/strong><\/td>\nControl Group<\/strong><\/td>\nP value<\/strong><\/td>\n<\/tr>\n
<\/td>\nNumber<\/td>\n%<\/td>\nNumber<\/td>\n%<\/td>\n<\/tr>\n
CMV CISH Positive Score<\/td>\nNegative<\/td>\n28.00<\/td>\n70.00<\/td>\n34.00<\/td>\n85.00<\/td>\n0.108<\/td>\n<\/tr>\n
Positive<\/td>\n12.00<\/td>\n30.00<\/td>\n6.00<\/td>\n15.00<\/td>\n<\/td>\n<\/tr>\n
 <\/p>\n

 <\/p>\n

 <\/p>\n

CMV CISH Positive Intensity<\/p>\n

 <\/td>\n

Score\u00a0 I<\/td>\n6.00<\/td>\n50.00<\/td>\n5.00<\/td>\n83.30<\/td>\n0.308<\/td>\n<\/tr>\n
Score\u00a0 II<\/td>\n3.00<\/td>\n25.00<\/td>\n1.00<\/td>\n16.70<\/td>\n<\/td>\n<\/tr>\n
Score\u00a0 III<\/td>\n3.00<\/td>\n25.00<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
weak\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 \/ I<\/td>\n7.00<\/td>\n58.30<\/td>\n6.00<\/td>\n100.00<\/td>\n<\/td>\n<\/tr>\n
Moderate \/II<\/td>\n5.00<\/td>\n41.70<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
strong\u00a0\u00a0\u00a0\u00a0\u00a0 \/ III<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

The Validity of CISH and IHC techniques for diagnosing placental tissues of miscarriage patient infected with EBV showed that of the total 40 placental tissues in the miscarriage group, nine have been identified to express CISH reaction for EBERs and IHC reaction VCA. No numerically important differences between these results of the two detection methods (P<0.005). The statistical observed sensitivity and specificity were\u00a0 44.4% and 83.9%, for CISH and IHC for detection of EBV in placental tissues of miscarriage patients respectively (Table 5), while the validity of CISH and IHC techniques\u00a0 in\u00a0 diagnosis CMV placental infections showed that the percentage of positive \u2013reaction results of\u00a0 CMV \u2013DNA -CISH in placental tissues of miscarriage patients was 30%\u00a0 while\u00a0 CMV-IHC\u00a0 protein expression\u00a0 in placental tissues of miscarriage\u00a0 group was 37.5%\u00a0 .\u00a0 The number of cells in serial sections that showed positive IHC was slightly greater than those have showed CISH positive reaction. <\/strong>Labeling results of both in situ hybridization and immunohistochemistry were showed in five cases. Statistically the observed sensitivity and specificity were 41.7% and 64.3%, for CISH and IHC for detection of CMV in placental tissues of miscarriage patients, respectively (Table 6).<\/p>\n

Table 5:\u00a0 Validity of EBV- CISH and EBV- IHC <\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\u00a0<\/strong><\/p>\n

\u00a0<\/strong><\/p>\n

EBV IHC Score<\/strong><\/td>\n

Miscarriage Group<\/strong><\/td>\n<\/tr>\n
EBV CISH<\/strong><\/td>\n<\/tr>\n
Negative\u00a0<\/strong>(n=31)<\/strong><\/td>\nPositive\u00a0<\/strong>(n=9)<\/strong><\/td>\n<\/tr>\n
No<\/td>\n%<\/td>\nNo<\/td>\n%<\/td>\n<\/tr>\n
Negative<\/td>\n26.00<\/td>\n83.90<\/td>\n5.00<\/td>\n55.60<\/td>\n<\/tr>\n
Positive<\/td>\n5.00<\/td>\n16.10<\/td>\n4.00<\/td>\n44.40<\/td>\n<\/tr>\n
P value<\/td>\n<\/td>\n0.073<\/td>\n<\/td>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Table 6: Validation of CMV for DNA CISH and for CMV protein IHC among the placental miscarriage tissues.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\u00a0<\/strong><\/p>\n

\u00a0<\/strong><\/p>\n

CMV IHC Score<\/strong><\/td>\n

Miscarriage Group<\/strong><\/td>\n<\/tr>\n
CMV CISH Score<\/strong><\/td>\n<\/tr>\n
Negative (n=28)<\/strong><\/td>\nPositive<\/strong><\/p>\n

(n=12)<\/strong><\/td>\n<\/tr>\n

No<\/td>\n%<\/td>\nNo<\/td>\n%<\/td>\n<\/tr>\n
Negative<\/td>\n18<\/td>\n64.30<\/td>\n7.00<\/td>\n58.30<\/td>\n<\/tr>\n
Positive<\/td>\n10<\/td>\n35.70<\/td>\n5.00<\/td>\n41.70<\/td>\n<\/tr>\n
P value<\/td>\n<\/td>\n0.722<\/td>\n<\/td>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Discussion<\/strong><\/p>\n

EBV could be the reason of infection to placenta in pregnancy, with consequent complications fetus. Primary and secondary EBV infections may both occur during pregnancy. However, main infections with EBV seems trans placental diffusion were occasional, whereas secondary maternal EBV infections weren\u2019t rare (Avgil and Ornog\u00a0 2006).<\/p>\n

Most of laboratory tests that were performed to diagnose EBV infections (including infections during pregnancy) use molecular techniques (like PCR) and serological tests for EBV antigens or antibodies specific to EBV by ELISA, immunofluorescent technique, or rapid tests (<\/em>Christian et al.<\/em>,<\/em> 2012).\u00a0 In this study, 9 out of 40 (22.5%) of the examined placental tissues were positive for EBV (EBER) by CISH\u00a0\u00a0 technique. Also the appearances of EBV-VCA were distinguished at nuclear localization in 22.50% (9 out of 40). In the current study which is\u00a0 presented here,\u00a0 a partial measurable CISH method was used for\u00a0 scoring of bleach growth,\u00a0 according to the number of placental cells infected with EBV and the results showed that most of infections have\u00a0 of score I, and this\u00a0 might represents the comparative mild to reasonable EBV infections of placental tissue through pregnancy as well as these findings were\u00a0 supported by measurement of color intensity of positive cases were most of the examined\u00a0 slides were having mild signal\u00a0 intensity.<\/p>\n

In the present study, the positive placental tissues of miscarriage group may indicate recently infected, or lytically reactivated EBV infection, however, the presence of EBER as detected by CISH, does not always correspond with the presence of VCA by IHC.<\/strong><\/p>\n

Immunohistochemistry was applied for detection VCA of EBV in placental tissue samples and it showed compatibility of those obtained by ISH and this might indicate a high compassion and specially the IHC-VCA\u00a0\u00a0 test, since the EBER in situ<\/em> hybridization for<\/em> detecting of latent EBV infection is considered the gold standard method and as supported by the finding of Truong et. al<\/em>., 2009. IHC has also been used where\u00a0 in this regard , \u00a0Xiao and his worker, 2014 have \u00a0showed\u00a0 also\u00a0 strong agreement for both techniques which\u00a0 were used\u00a0 for detection EBV in patients with lupus nephritis.<\/p>\n

The present result are supported by Xuan-Hong (2011) who\u00a0 suggested\u00a0 that the\u00a0 possibility of the EBV virus to\u00a0 transmission\u00a0 from the\u00a0 uterus to the fetus , which were\u00a0 resulting in stillbirth , abortion , or congenital defects.<\/p>\n

The current study provide a novel information about the histological localization of EBV nucleic acid in placental tissues using CISH, were\u00a0 the overall frequency of EBV infection is 22.5 % of the examined placentae, the present result\u00a0 is higher than those reported by other abroad studies such as that\u00a0 done by Gervasi and his colleagues (Gervasi et al<\/em>.,2012 ) who found\u00a0 viral genomic\u00a0 sequences of EBV (0.1%) of their examined cases of\u00a0 the amniotic fluid\u00a0(\u00a0in the mid trimester of pregnancy) \u00a0by using quantitative real-time PCR for the presence of EBV-genome sequences. Recent studies done by Song and his Colleagues in 2015, used ISH for detection EBV in lympho-proliferative disorders, nasopharyngeal carcinoma and related malignancies with final color or fluorescence detection however, there is no reported data on detection of EBV by ISH during pregnancy and this signify a novelty for this study(Song, 2015 ).<\/p>\n

Human cytomegalovirus is an significant etiological agent of intrauterine infection, which may lead to some thoughtful consequences in pregnant women such as miscarriage, stillbirth, cerebellar malformation and fetus development retardation, as depicted in Staar et., al<\/em> 2012.<\/p>\n

The results of this study was in line with a serological study done in Iraq to discover the association of TORCH infections\u00a0 in women with spontaneous abortions , CMV Ab prevalence\u00a0 of\u00a0 positive\u00a0 cytomegalovirus\u00a0 CMV(38.50%) in 2009 and ,\u00a0 the lowest percentage were observed\u00a0 only\u00a0 (29.10%) of cases\u00a0 in 2010. This could suggest that the exposure to CMV infection was declined over this period in Iraq. \u00a0<\/strong>\u00a0(Majeed, 2011)<\/p>\n

The study that was achieved in Baghdad by (Maysara et., al <\/em>2012)\u00a0 to evaluate the prevalence of seropositivity of specific IgM antibody for CMV by ELISA, CMV specific IgM antibody was detected in 15.7% of the 108 women with history of abortion.<\/p>\n

In Australia, (Jenna et., al<\/em> 2015) conclude to \u00a0the result that the total CMV DNA was distinguished in 5.0 % of\u00a0 placenta soft tissue of miscarriage women along with the\u00a0 infections were confirmed by using the\u00a0 immune-histochemical assay and viral\u00a0 proteins.<\/p>\n

Additional seroepidemiological research was made in Havana which was made by (Aim\u00e9e et., al<\/em> 2015) at maternity hospital by means of marketable ELISA kits has shown that the prevalence of active cytomegalovirus infection was detected in 16.70 %,.<\/p>\n

Our results disagree using a\u00a0 previous outcomes\u00a0 done by \u00a0Sharief,( 2005) , who showed an absolute positive\u00a0 results of aborted women with a\u00a0 primary abortion i.e\u00a0 reveled\u00a0 highest percentage (100% )\u00a0 which was\u00a0 considered\u00a0 to carry a\u00a0 high titer of IgG. In developed countries such as\u00a0 United States it was\u00a0 stated that HCMV infection at rate of\u00a0 80% was observed\u00a0 at age 35-40 years old women who\u00a0 have anti- HCMV IgG\u00a0 in comparison to\u00a0 50% -80% HCMV infection in the\u00a0 younger age\u00a0 women\u00a0 (Gold, and Nankervis, 2007).\u00a0 The hematogenous way of CMV diffusion in the placenta can clarify the principal infection to this microbe in the variable villi.<\/p>\n

The current study revealed that sensitivity of IHC is slightly higher than that of ISH for histological detection of CMV, this might be due to CMV protein over expression (which was studie here) can be detected by IHC but not by ISH, similar result were also obtained by LU DY and his colleagues (2009) found similar findings that the result of IHC is higher than those of ISH; in our opinion the CISH is more difficult to done and this might slightly affect the sensitivity of the procedure.<\/p>\n

Yiska et., al <\/em>(2011) did an Immunohistochemical analysis to infected units to \u00a0parental decidua exposed the appearance of CMV immediate-early and pp65 early-late virus-related genetic factor and gB as well which stated late later infection. The control tissues were found negative by immunohistochemical staining. These findings indicate that HCMV, in the infected placental tissues undergoes a full replication cycle.<\/p>\n

While this research dominant proportion of control group (6 from 40 using CISH) and (2 from 40 using IHC), this reflect to CMV positivity in these placental soft tissues in controller groups may reflect the need for following the studies values of the infections of such pregnancies both on toddlers and mothers subsequently later the deliveries cause the abortion consequences of placental CMV infection thru pregnancy and postnatal consequence should be well thought-out as well.<\/p>\n

The present research follow\u00a0 the inclusion patients with abortions, and omitted the other results of\u00a0 infected pregnancy like fetal inherited anomalies and illnesses, it represent a restriction to this study which was established by recording as well as the difficult understanding of CMV infection in the control placental tissue\u00a0 group.<\/p>\n

In study in Iraq , Alwan and\u00a0 Sera\u00a0 (2011) have\u00a0 indicated\u00a0 that CMV infection rate was (46.6%),These results were\u00a0 higher than our results ,\u00a0 this different in the\u00a0 CMV prevalence rate \u00a0may be due to different in the sample\u00a0 size included in the study group\u00a0\u00a0 as well as may be due\u00a0 to substantial diffusion of infection in local population , perhaps\u00a0\u00a0\u00a0 that CMV\u00a0 is endemic in Iraq during that period ,\u00a0 or may be CMV infections\u00a0 are un common in a particular region\u00a0 .<\/p>\n

The current results obtained from\u00a0 both techniques\u00a0\u00a0 for EBV have revealed that among\u00a0\u00a0 Forty\u00a0 placental tissues\u00a0 from\u00a0 miscarriage patients ,\u00a0\u00a0 EBER- CISH and VCA-IHC have been found in 22.5% . While\u00a0 the\u00a0 detection rates of EBV-CISH and EBV-IHC in placental tissue sections were equivalent\u00a0 yet the validity results\u00a0 EBER by ISH where compared to\u00a0 VCA by IHC\u00a0 had displayed a sensitivity and specificity of 44.40 % and 83.90 %, respectively,<\/p>\n

Also in this study the evaluation of the efficacy of these two procedures but for detection the presence of CMV infection in placental tissues from miscarriage patients. Both CISH and IHC for diagnosing\u00a0 nuclear inclusions\u00a0 of\u00a0 this virus in placental tissues have showed that , CMV-DNA- CISH and CMV-protein -IHC were found in 30% , 37.5 %\u00a0 respectivly. The comparism\u00a0 of\u00a0 the results of\u00a0 both technique\u00a0 revealed\u00a0 a sensitivity and specificity of 41.7% and 64.3%, respectively. The numbers of infected cells scored were slightly higher with IHC than with in CISH technique. These results show that the use of both in CISH and IHC for EBERs and the EBV- VCA as well as CMV \u2013DNA and CMV proteins are both specific, as well as sensitive methods.<\/p>\n

The current result of this study are matching with Niedobitek et<\/em> al<\/em>.,(1988) result . They used ISH, IHC, and morphological analysis of tissue from patients with AIDS who have widespread CMV infections.\u00a0 They showed that the evaluation of the results in ISH and IHC were significantly extra sensitive than the morphological analysis.\u00a0 It was further shown that IHC has shows a higher CMV infection in cells than in CISH. They concluded that IHC as technique appeared to be more suitable to be used.<\/p>\n

Several authors have denoted that ISH was considered as a method of optimal to detect EBERs \u2013EBV presence inside tissues section and this is because of\u00a0 the huge data of duplicates of EBERs present in infected cell where\u00a0 these works have\u00a0 showed Positive staining in the nuclei of the EBV-infected cells (Weiss and Chen , 2013;\u00a0 H\u00e4nel et.al., 2001; Margaret \u00a0and Weihua , 2008 ).<\/p>\n

In addition our data were match with the fact that ISH was considered as the gold standard for precisely identifing the sites of expression of EBV as it and was highly sensitive and specific; however, some authors denoted that ISH is expensive (Lerner et al<\/em>. ,1981). The results of the current study showed that the two detection techniques had strong agreement, which in turn indicated that the results of both were reliable.<\/p>\n

In conclusion CMV and EBV are important causes of placental infections among miscarriage females in Baghdad, and CMV might be detected in placenta of normal delivery. Although \u00a0CISH<\/em> technique considered as the gold standard method for<\/em> detecting of\u00a0 latent EBV and \/or CMV infection were IHC has showed a\u00a0 compatibility\u00a0 to\u00a0 that technique\u00a0 and might reach rates of\u00a0 high sensitivity and specificity similar to it.<\/p>\n

\u00a0<\/strong>References<\/strong><\/p>\n

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