The incidence of cytomegalovirus (CMV) antigenemia and CMV disease is reduced by highly active antiretroviral therapy.

Cytomegalovirus (CMV) infection was one of the most common opportunistic infections in AIDS patients, leading to blindness or life-threatening disease in about 40% of patients in the later stages of AIDS before highly active antiretroviral therapy (HAART). In a retrospective multicenter study we investigated the incidence of CMV retinitis and organ involvement in Northern Italy before (1995 and 1996) and after the introduction of HAART (1997 and 1998) as well as the data regarding CMV antigenemia. We found a sharp drop in the incidence of CMV disease in AIDS patients as well as a decline in the incidence of relapses of CMV-disease after the widespread introduction of HAART. Moreover, there was a decrease in the incidence of antigenemia-positive cases in AIDS patients in the era of HAART and the median CMV viral load was significantly higher in patients who didn't receive HAART than in patients who received HAART (p = 0.001, t test).

Maternal IgG avidity and IgM detected by blot as diagnostic tools to identify pregnant women at risk of transmitting cytomegalovirus.

In this study, we determined the avidity index (AI) of anticytomegalovirus (CMV) immunoglobulin G (IgG) and the anti-CMV immunoglobulin M (IgM) profile in 124 pregnant women, 87 of whom were considered at risk of transmitting CMV infection to their offspring and 37 of whom were at no risk. IgG avidity and blot for IgM were performed on two serum samples from each woman, at 6-18 weeks' gestation and at 20-23 weeks' gestation. Pregnancy outcomes were monitored. The results obtained showed that the determination of anti-CMV IgG avidity at 6-18 weeks' gestation can identify all women who would have an infected fetus/newborn (100% sensitivity), whereas IgM detected by blot had poorer results (69% sensitivity). Interestingly, at 20-23 weeks' gestation, the sensitivity of IgM detection by blot was higher than that obtained by avidity (75 % and 63%, respectively) and the combination of IgG avidity and IgM by blot yielded the best results (81% sensitivity).

Anticytomegalovirus (anti-CMV) immunoglobulin G avidity in identification of pregnant women at risk of transmitting congenital CMV infection.

In this work, we show that the determination of the anticytomegalovirus antibody avidity carried out before week 18 of gestation is a helpful tool to identify women for enrollment in prenatal diagnosis. This procedure can identify all pregnant women who will give birth to an infected newborn.

New advances in the diagnosis of congenital cytomegalovirus infection.

With the advances in anticytomegalovirus (anti-CMV) serology, the new recombinant IgM tests seem likely to become the screening tests for pregnant women whose prepregnancy serological status for CMV is unknown. When a woman is found to be IgM-positive, further diagnostic evaluation focused on determining whether this is due to a primary infection should be carried out. Maternal primary infections that were difficult to determine until a few years ago unless documented by seroconversion can now be readily diagnosed from the presence of low-avidity anti-CMV antibody which persists for approximately 20 weeks after primary infection. In primarily infected mothers prenatal diagnosis can be performed between 21 and 23 weeks of gestation, and the amniotic fluid (AF) represents the pathological material of choice to determine intrauterine virus transmission. In AF, the virus can be detected by culture and/or PCR. Both procedures differentiate uninfected from infected fetuses, but cannot predict fetal outcome. The determination of the viral load in AF carried out by quantitative PCR is more promising and could represent an important starting point for preemptive fetal therapy.

Prenatal diagnosis of congenital cytomegalovirus infection.

We report here the results of a study on the prenatal diagnosis of congenital cytomegalovirus (CMV) infection. The study was carried out by both PCR and virus isolation from amniotic fluid (AF) for 82 pregnant women at risk of transmitting CMV for the detection of (i) seroconversion to CMV immunoglobulin G (IgG) positivity during the first trimester of pregnancy, (ii) symptomatic CMV infection in the mother during the first trimester of pregnancy or intrauterine growth retardation detected by ultrasound or abnormal ultrasonographic findings suggestive of fetal infections, and (iii) seropositivity for CMV-specific IgM. For 50 women, fetal blood (FB) was also obtained and tests for antigenemia and PCR were performed. The results indicate that AF is better than FB for the prenatal diagnosis of CMV infection. PCR with AF has a sensitivity (SNS) of 100%, a specificity (SPE) of 83.3%, a positive predictive value (PPV) of 40%, and a negative predictive value (NPV) of 100%; rapid virus isolation with the same material has an SNS of 50%, an SPE of 100%, a PPV of 100%, and an NPV of 94.7%. Fewer than 10% of the women positive for IgM by enzyme immunoassay (EIA) had a congenitally infected fetus or newborn infant. When EIA IgM positivity was confirmed by Western blotting (WB) and the WB profile was considered, the percent transmission detected among women with an "at-risk" profile was higher than that observed among IgM-positive women and was the same as that among women who seroconverted during the first trimester of pregnancy (transmission rates of 29 and 25%, respectively).

Delayed acquisition of high-avidity anti-cytomegalovirus antibody is correlated with prolonged antigenemia in solid organ transplant recipients.

Previous studies have demonstrated that maturation of cytomegalovirus (CMV)-specific antibodies in solid organ transplant recipients is delayed after primary CMV infection. To investigate the clinical significance of this finding, the avidity indices of anti-CMV antibody were determined in parallel with other serologic and virologic parameters in serial serum samples from 24 solid organ transplant recipients who had primary CMV infection that began during the first 3 months after transplantation. The data obtained show that a delay in antibody maturation is significantly correlated with a long persistence of positive antigenemia.

Development of a new cytomegalovirus (CMV) immunoglobulin M (IgM) immunoblot for detection of CMV-specific IgM.

We developed a new cytomegalovirus (CMV) immunoglobulin M (IgM) immunoblot to detect CMV-specific IgM in human sera. The new test contains four viral proteins (vp150, vp82, vp65, and vp28) purified from viral particles and four recombinant proteins (rp150, rp130, rp52, and rp38) purified from Escherichia coli. These antigens were individually loaded onto nitrocellulose strips, and the strips were then used to detect CMV-specific IgM by using a mu-specific conjugate. The new assay was evaluated in parallel with one or two IgM enzyme-linked immunosorbent assays (ELISAs) to test 592 serum samples from different groups of latently or acutely infected individuals. The sensitivity of the new assay with respect to the consensus of two ELISAs was 100%, the specificity was 98.6%, the positive predictive value was 96.9%, and the negative predictive value was 100%. We also evaluated the new test by testing sera from pregnant women and transplant recipients with a known clinical history. Our results suggest that the new test combines high sensitivity with high specificity, characteristics that are mutually exclusive with the other commercially available tests. Furthermore, a statistically significant correlation was observed between the number of IgM-reactive bands and the elevated risk of transmission from CMV-infected pregnant women to their offspring.

Cytomegalovirus infection in pregnancy: a still complicated diagnostic problem.

The diagnostic problems linked to human cytomegalovirus (HCMV) in pregnancy are many and not all have been fully defined. In long-term seropositive women there is a tacit agreement that no laboratory testing for HCMV should be carried out. In seronegative women a test for HCMV-specific IgG should be performed at least twice during the first 4 months of pregnancy, and if the seronegativity persists, further follow-up might be stopped. On the other hand, if a seropositivity appears the diagnosis of a primary HCMV infection is established and prenatal diagnosis should be offered to the mother. Finally, in the case of a pregnant woman with unknown serological status, the diagnosis of HCMV infection is a complex problem and several different questions need to be addressed. In our opinion they should be screened with a reliable IgM test (confirmed by blot if necessary) followed, in the case of positivity, by an avidity assay. Pregnant women undergoing a primary HCMV infection should be encouraged to seek prenatal diagnosis to be performed by PCR and virus isolation from amniotic fluid at the 21st to 23rd week of gestation.

Quantitative cytomegalovirus (CMV) antigenaemia during antiviral treatment of AIDS-related CMV disease.

In order to assess the value of quantitative measurement of cytomegalovirus (CMV) antigenaemia as a marker for the guidance of antiviral chemotherapy in the AIDS setting, 33 patients with CMV complications and showing at least 20 pp65-positive polymorphonuclear leucocytes per 2 x 10(5) cells, received either ganciclovir or foscarnet as induction and maintenance therapy. Antigenaemia was assessed every 1-4 weeks. During acute-phase antiviral therapy, a significant decrease of CMV antigenaemia (>50% of pretreatment levels) paralleled clinical improvement in 2-7 weeks in 32 of 33 subjects. In ten of 24 evaluable patients followed up during a further 4-12 months, disease relapses occurred concurrently with an increase of CMV antigenaemia in seven cases, while three cases of relapsing retinitis did not show a significant increase in antigenaemia. All patients with recurrent disease had a favourable response to further treatment, including halted clinical progression and significant decrease in antigenaemia. In HIV-related CMV disease, periodic monitoring of quantitative CMV antigenaemia proves useful in evaluating response to antivirals, in guiding therapeutic management and in predicting disease relapses.

Avidity of immunoglobulin G directed against human cytomegalovirus during primary and secondary infections in immunocompetent and immunocompromised subjects.

Diagnosis of primary human cytomegalovirus (HCMV) infection is accomplished exclusively by serologic testing. Among the possible methods, the determination of immunoglobulin G (IgG) avidity is one of the least explored. In this work, we used a commercially available kit to test anti-HCMV IgG avidity in 336 serum samples from pregnant women and transplant recipients undergoing virologically proven HCMV primary or nonprimary infections and from latently infected blood donors. Our results demonstrate that the anti-HCMV IgG avidity test differentiates primary from nonprimary HCMV infections in both pregnant women and solid organ transplant recipients. In fact, 88.6% of primary infections and no secondary infections showed low-avidity IgG to HCMV. In particular, low IgG avidity is a marker of primary infection for 18 to 20 weeks after onset of symptoms in both immunocompromised and immunocompetent subjects.

A quantitative test (HCMV-hybrid-capture(TM)) to detect human cytomegalovirus DNA in the blood of immunocompromised patients compared with antigenemia and polymerase chain reaction.

A quantitative assay is necessary to differentiate between low and high Cytomegalovirus load in the blood, a high load frequently being correlated with clinical disease. A new method for the quantitative determination of viral DNA (HCMV-hybrid-capture) was compared with antigenemia and PCR. Hybrid-capture proved to be a simple and rapid method for the quantitative determination of viral load in the blood. It is less sensitive than PCR and antigenemia, but seems to correlate with clinical symptoms as well as the antigenemia test and better than non quantitative PCR.