Real-life evaluation of a human immunodeficiency virus screening algorithm using a single combined p24 antigen-antibody assay.

Since May 2010, human immunodeficiency virus (HIV) screening in France has been performed using a single combined fourth-generation assay. One of our major concerns is to verify that this screening strategy is able to diagnose HIV primary infection as soon as possible. Thus, the sensitivity and specificity of this strategy were evaluated on 49,623 serum samples, including 29 primary infections, received for routine HIV testing between September 2010 and November 2011. Specimens were screened using the Enzygnost HIV Integral II enzyme-linked immunosorbent assay (ELISA) kit. All positive sera, according to the manufacturer's recommendations [signal-to-cutoff ratio (S/CO) ≥ 1] were retested using the Architect HIV Ag/Ab Combo. Moreover, we defined a grey zone (0.5 < S/CO < 1) and sera within this grey zone were retested using the VIDAS HIV DUO Ultra test and HIV-1 RNA was checked by the Abbott RealTime PCR kit. Screening tests were positive for all primary infections. All samples within the grey zone proved VIDAS HIV DUO Ultra and HIV-1 RNA negative. Overall, the ELISA test sensitivity and specificity were 100 and 99.79 %, respectively. The false-positive rate was higher when S/CO was in the low range (1 to 5). Adding a second screening test for positive sera reduced the false-positive rate from 0.20 to 0.02 %. HIV screening with a single combined assay did not miss any documented primary infection during this evaluation period, even without extending the positivity zone.

Prospective evaluation of the threat related to the use of seminal fractions from hepatitis C virus-infected men in assisted reproductive techniques.

BACKGROUND: The risk of hepatitis C virus (HCV) transmission during assisted reproductive techniques (ARTs) is still disputed and no report concerning its prospective evaluation is available. METHODS: The aim of this 4-year follow-up multicentre study that enrolled 86 HCV-serodiscordant couples was to determine whether a sperm-processing method was able to reduce levels of HCV in semen and the risk of HCV transmission to the newborn. All the men were chronically infected by HCV and 10 of them by human immunodeficiency virus. A total of 181 seminal plasmas and 153 sperm fractions were tested for the presence of HCV RNA. RESULTS: HCV RNA tested positive in 20.4% of the seminal samples. All of the 153 final sperm fractions tested negative for HCV. The detection of HCV RNA in semen was significantly correlated with a high viral load in blood (P < 0.05). The presence of HCV RNA in seminal plasma impaired neither semen parameters nor ART issue. From the 58 couples enrolled effectively in an ART programme, 24 pregnancies and 28 newborns were obtained. All of them tested negative for HCV RNA in blood. CONCLUSION: These results emphasize the safety of the semen-processing method. The negligible risk of transmitting HCV reduces the value of the systematic analysis of HCV RNA in seminal fractions prior to ART. Since use of this analytical procedure involves the freezing of semen, its avoidance would result in an increase in sperm quality and reduce the need to perform intracytoplasmic sperm injection techniques.

[Redox status in HIV+ patients under HAART]. / Statut oxydatif de patients VIH positifs sous trithérapie.

Oxidative stress decreases immune defences and is also suggested to participate in the activation of HIV virus replication. That is why we decided to explore some biomarkers of oxidative stress (reduced glutathione, lipoperoxides, true malondialdehyde and vitamin C) in 20 HIV positive patients whose HIV replication was determined by measurement of RNA viral load. Reduced glutathione is decreased in HIV positive patients, without correlation with the viral load. The patients mean content of lipoperoxides is twice that of controls but with such a large range that there is no statistical difference.

Transmission risk of hepatitis C virus in assisted reproductive techniques.

Medical assistance for procreation in a couple where one or both parents has hepatitis C viral infection (HCV) raises the issue of the transmission of the infection to the baby and/or of possible contamination of both the technicians and the gametes or embryos from virus-free parents in the laboratory. It becomes essential to assess transmission risk in assisted reproductive techniques in order to define clearly the management of couples according to their viral status. To define the HCV transmissibility risk in assisted reproduction related to the presence of virus in semen from infected infertile men, HCV RNA detection was performed in sera, and semen and sperm fractions obtained after Percoll gradient centrifugation. HCV RNA was detected in 5% (2/39) of the semen samples tested: in the raw semen, in the seminal fluid and in the cell pellet but never after Percoll selection. According to these results, we suggest a strategy for HCV-infected infertile men who need assistance for procreation.

Gene transfer of anti-gp41 antibody and CD4 immunoadhesin strongly reduces the HIV-1 load in humanized severe combined immunodeficient mice.

OBJECTIVE: To study the anti-HIV-1 effects of the delivery of anti-gp41 monoclonal antibody (mAb) and soluble CD4 (sCD4) immunoadhesin by genetically modified cells in HIV-1-infected, humanized severe combined immunodeficient (SCID) mice. DESIGN: The complementary DNA of mAb 2F5, an anti-HIV-1 gp41 antibody, and of sCD4-IgG chimeric immunoadhesin were transferred into 3T3 cells using Moloney murine leukaemia virus vectors. The cells were then incorporated into a collagen structure called the neo-organ, which allowed the continuous production of the therapeutic molecules. METHODS: The antiviral effects in vivo of 2F5 or sCD4-IgG or both compounds were evaluated in neo-organ-implanted SCID mice that were grafted with human CD4 CEM T cells and challenged with HIV-1 Lai or MN. RESULTS: In SCID mice implanted with 2F5 neo-organs, antibody plasma levels reached 500-2000 ng/ml. Viral loads after HIV-1 challenge were significantly reduced in neo-organ-implanted HIV-infected mice. Although 29 x 10(7) and 13 x 10(8) HIV-1-RNA copies/ml were detected at 12 days in the controls (mice injected with Lai and MN, respectively) less than 16.5 x 10(3) HIV-1-RNA copies/ml were observed in all implanted mice injected with either Lai or MN. The intracellular viral load was also reduced in CD4 cells recovered from the implanted mice. Comparable antiviral effects were obtained with CD4-IgG neo-organs. CONCLUSION: Our results confirm the anti-HIV properties of 2F5 and sCD4-IgG continuously produced in vivo after ex-vivo gene therapy in SCID mice.

Experimental gene therapy: the transfer of Tat-inducible interferon genes protects human cells against HIV-1 challenge in vitro and in vivo in severe combined immunodeficient mice.

OBJECTIVES: To evaluate in vitro and in vivo a strategy for gene therapy for AIDS based on the transfer on interferon (IFN)-alpha, -beta and -gamma genes to human cells. DESIGN: Human U937 promonocytic cells were stably transfected with Tat-inducible IFN expression vectors conferring an antiviral state against infection with HIV. METHODS: Transfected cells were either infected by HIV-1 in vitro or transplanted into severe combined immunodeficient (SCID) mice for an HIV challenge in vivo. RESULTS: U937 cell lines stably carrying IFN transgenes under the positive control of the HIV-1 Tat protein were highly resistant to HIV-1 replication in vitro. This antiviral resistance was associated with a strong induction of IFN synthesis immediately following the viral infection. HIV-1 proteins were found to be specifically trapped within the genetically modified cells. In contrast, all IFN-U937 cells permitted full HIV-2 replication. Transfected cells injected into SCID mice and challenged against HIV-1 were strongly resistant to infection when cells were transduced with IFN-alpha of IFN-beta genes. However, IFN-gamma-transfected cells permitted HIV-1 infection in vivo despite the induction of a high level of IFN-gamma secretion. The quantity of proviral DNA was 10(5)-fold lower in IFN-alpha- or IFN-beta-transfected U937 cells collected from these SCID mice than that in non-transfected cells. CONCLUSIONS: Our results substantiated the validity of a strategy, bases on the transfer of HIV-1-inducible IFN-alpha or IFN-beta genes, to confer antiviral resistance to human cells.

Co-infection by hepatitis B virus and hepatitis C virus in renal transplantation: morbidity and mortality in 1098 patients.

The aim of the study was to analyse the influence of co-infection by hepatitis B virus (HBV) and hepatitis C virus (HCV) as compared with HCV infection alone in 1098 patients who received a kidney transplant between 1 January and 31 December 1991. At transplantation, the prevalence of anti-HCV antibodies was 21.40% (235/1098) while the prevalence of HBV infection was 9.85% (108/1096); 46 patients were co-infected with HBV and HCV, either 19.70% of HCV-infected patients and 42.60% of HBV-infected patients. Liver tests, galactose clearance and liver biopsy were compared in the 46 co-infected patients (HCV+HBV+) and in the 189 HCV-infected patients (HCV+HBV-). At the time of transplantation, cytolysis was present in 31.45% of HCV+HBV- patients (50/159) and in 40% of HCV+HBV- patients (16/40); cholestasis was present in 34.18% of HCV+HBV- patients (34/158) and 42.11% of HCV+HBV+ patients (16/38). At 6 months the incidence of biological abnormalities increased to 37% in HCV+HBV- patients (55/150) and to 52.5% in HCV+HBV+ patients (21/40), suggesting a more deleterious effect of the immunosuppressive therapy in the co-infected group. Over the course of transplantation, chronic hepatitis was present in 50% of HCV+HBV- patients and in 64.1% of HCV+HBV+ patients. Liver failure occurred in 7% of HCV+HBV- patients (12/156) and 17% of HCV+HBV+ patients (7/41). Galactose clearance was performed as a functional test in 68 patients: it was not significantly different in either group. Liver biopsy was performed in 108 patients at least once.(ABSTRACT TRUNCATED AT 250 WORDS)

Etude de la prevalence des anticorps anti-CMV seriques parmi la population congolaise seronegative et seropositive pour le VIH 1

L'objet de cette etude est d'apprecier la seroprevalence des anticorps de classe IgG et IgM dirigee contre le CMV parmi la population generale congolaise et de la comparer a celle observee chez les patients febriles seropositifs pour le VIH 1. Les bilans bacteriologiques; mycologiques et parasitaires sont negatifs. Les serums proviennent de 100 patients seropositifs pour le VIH 1 en ELISA; confirmes en WESTERN-BLOT et d'un groupe temoin de 100 sujets seronegatifs pour le VIH 1 en ELISA. [abstract terminated]

Cytomegalovirus infection--an etiological factor for rejection? A prospective study in 242 renal transplant patients.

The study aimed at analyzing the role of CMV infection as a risk factor for rejection occurring after CMV infection because of the clinical consequences of the prevention of CMV infection that might lead to the decrease in rejection episodes. Two hundred forty-two consecutive renal transplant patients were prospectively checked for the occurrence of CMV infection. CMV infection was defined virologically by a positive viremia or/and a positive viruria or/and a seroconversion or/and a significant rise of the anti-CMV antibody titers. Viremia, viruria, and serology were performed weekly for the first month and then at day 90, day 180, and every 6 months, and moreover if clinical symptoms related to a viral infection occurred. Rejection episode was defined by a creatininemia rise of 25%, after cyclosporine nephrotoxicity and urological complications had been discarded, and by the response to the antirejection therapy, steroids, or OKT3 in case of steroid-resistant rejection. The outcome factor was rejection episode occurring from day 4 after the diagnosis of CMV infection. A patient undergoing "a rejection episode after CMV infection" could also be exposed to other potential confounding factors that can be considered as risk factors of rejection among our patients. Rejection occurring before CMV infection was the main factor because it was linked both to CMV infection itself and to "rejection after." Thus infected and noninfected patients were randomly paired off. To the noninfected patient of the pair was attributed the date of a fictitious CMV infection that was the date of the CMV infection of the infected member of the pair. Therefore, "rejection after" and "rejection before" were defined in infected and noninfected patients of the pair according to the time of onset of CMV infection of the infected member of the pair. The incidence of CMV infection was 65%, 157 of the 242 patients were infected, and 85 not infected. Thus 85 pairs of infected-noninfected patients were studied. The incidence of "rejection after" the diagnosis of CMV infection was significantly higher in the group of patients with CMV infection: 45% among infected (38/85) versus 10.60% among noninfected (9/85) (P < 0.0001). Among the 85 pairs, 48 pairs were concordant in which patient of the pair evinced the same outcome factor: 43 showed no rejection after, and 5 showed one.(ABSTRACT TRUNCATED AT 400 WORDS)

Cytomegalovirus (CMV) excretion as a factor in the severity of CMV disease in kidney and simultaneous kidney and pancreas transplantation.

The aim of the study was to evaluate the virological parameters associated with the severity of cytomegalovirus (CMV) disease in renal and simultaneous renal and pancreatic transplantation. The association of the viral profile and the severity of the viral disease was analysed taking into account different confounding variables susceptible to linkage with the severity of the CMV infection and the viral parameters. All the patients transplanted between 1 January 1989 and 31 December 1990, a total of 242, were prospectively followed by viral cultures in blood and urine and by serological methods using the detection of CMV-specific IgM and the complement fixation (CF) test. The samples were taken systematically each week for the first month and then at day 90, 180 and every 6 months and also in cases of clinical manifestations related to viral disease. CMV infection was diagnosed virologically by the presence of viraemia, viruria, IgM, or a significant rise in CMV antibody titre in CF. CMV disease was classified as asymptomatic, mild (fever and/or leukopenia), moderate (fever, leukopenia and liver abnormalities), severe (CMV pneumopathy and/or gastrointestinal disease) or fatal. The incidence of CMV infection was 65% (157/242): 32% asymptomatic, 36% mild, 30% moderate and 2% severe. The presence of IgM was associated with the severity of CMV disease: 51.4% of moderate and severe CMV infections in the group with IgM versus only 16% in the group without IgM (P < 0.0001). The risk of having severe or moderate CMV disease was 3.28 times higher in patients with positive IgM. However the serological changes in CF were not significantly associated with the severity of the viral disease since 34.6% of the patients with CF changes had a severe form versus 20.8% in the group without CF modification. Viruria was significantly associated with moderate or severe infection: 43.6% of the patients with viruria had severe infection versus only 12.5% in the patients without viruria (P < 0.0002). The risk of having moderate or severe CMV disease was 3.48 times higher in the patients with viruria. Viraemia was also associated with more severe CMV infection: 48.6% of moderate or severe CMV infection in the group of patients with viraemia versus 19% in the group without viraemia (P < 0.0001). The risk of having severe or moderate CMV infection was 2.58 times higher in the patients with viraemia. Viraemia was not more associated with severe CMV infection than viruria. Using the maximum likelihood ratio method and the logistic regression model, CMV-specific IgM, viruria and viraemia were each shown to be associated with the severity of CMV disease and the addition of one parameter to the other(s), whatever the type (except the CF changes) and whatever the order of this addition, did not remove the link between the severity and IgM, viruria and viremia. The incidence of severe and moderate CMV disease increased with the number of positive viral parameters (PVP) from 2% of moderate and severe infections in the group with one PVP, to 28% in the group with two PVP, to 39% in the group with three PVP and 68% in the group with four PVP (trend, 35.95; P < 0.0001). Taking the absolute risk of the group of patients without IgM, viruria or viraemia as the basal level, the observed relative risk of severe CMV infection varied from 6.45 in the group with positive IgM without viruria or viraemia, to 10.74 in the group with positive IgM and viruria without viraemia and to 22.5 in the group with the three positive parameters IgM, viruria and viraemia. The different potential confounding factors (recipient and donor serology, renal or renal and pancreatic transplantation, DR compatibility, rejection before CMV infection) did not modify the link between the viral profile and the severity of CMV disease. This study suggests that the severity of CMV disease might be linked to the overspread of the virus as well as to the consequences of a CMV-specific humoral immune response.

Should hepatitits-C virus antibody-positive donors be excluded from kidney donation?

In organ transplantation, virus transmitted by the donor is associated with a higher risk of severe primary infection after transplantation in the seronegative recipient. In this study, the risk of hepatitis-C virus (HCV) transmission by the kidney was determined, and the morbidity in the recipient assessed. Serum samples from all kidney donors of our Transplantation Unit between 1983 and 1988 were screened for antibodies to anti-HCV by first enzyme-linked immunosorbent assay (Ortho ELISA) and positive samples were confirmed by a second-generation ELISA and the CHIRON RIBA HCV test. Of the 164 kidney donors whose sera were available, five were positive (3%) and all of them were positive with the RIBA test. Liver function was normal in the five donors. Seven recipients received a renal transplant from the anti-HCV-positive donors. Two patients had a follow-up too short to draw any conclusions. Two patients remained anti-HCV-negative up to 36 and 48 months, respectively, but one of them had chronic hepatitis. One patient was anti-HCV-positive before transplantation and remained positive over the 4-year follow-up. The two last patients seroconverted and acute hepatitis occurred at 16 and 101 days after transplantation, respectively. In both cases, no peroperative or postoperative transfusion was given and no other cause of hepatitis could be determined. A cirrhotic evolution was observed within 15 and 36 months in both cases. Thus HCV can be transmitted by a kidney transplant and cadaveric donors positive for anti-HCV antibodies should be excluded from kidney donation.

Characterization of immune complexes containing cytomegalovirus-specific IgM antibodies following a kidney graft.

In order to demonstrate the viral specificity of IgM-containing immune complexes (IgM-CIC) detected by a C1q assay in renal allograft recipients developing a CMV infection, a technique is described allowing: 1) the dissociation of IgM-CIC by action of an acid buffer, and 2) the characterization of the viral specificity of IgM antibodies released by this treatment. This step was performed by ELISA and Western Blot. When technique was applied to the follow-up of a renal allograft recipient developing a recurrent CMV infection within 2 months post-graft, it was found that the IgM-CIC detected on the day of the graft were not CMV-specific, whereas the IgM-CIC detected during the second month after transplantation contained CMV-specific IgM antibodies. These CMV-specific IgM-CIC were detected as early as the urinary viral excretion. It was shown by Western Blot analysis that these IgM antibodies reacted with a 45-47 kDa viral polypeptide which is a viral target for specific humoral response at the early phase of CMV infection.

Acute meningoradiculitis concomitant with seroconversion to human immunodeficiency virus type 1.

We report a case of acute regressive meningoradiculitis concomitant with HIV1 primoinfection. The clinical symptoms were mild and disappeared spontaneously. Electromyographic studies confirmed the regressive demyelinating-type process. The biological diagnosis of HIV1 infection was demonstrated by viral antigen detection, by the presence of the integrated proviral DNA after gene amplification by a polymerase chain reaction, and in particular, by the steady progression in the appearance of different HIV1-specific antibodies. This was shown using three different Western blot kits whose performances were compared.

Blocking-ELISA for detection of specific antibodies to the glycoproteins of the human immunodeficiency virus type 1 (HIV-1).

A blocking ELISA was developed to confirm the specificity of screening tests for anti-HIV-1 antibodies. A murine monoclonal antibody (McAb) raised against recombinant gp160 was used in combination with a commercial technique (ELA-VIA-1). After determining the optimal experimental conditions, the assay was applied to 92 samples presenting different reactivities by Western blot (WB) analysis. All the sera containing antibodies to gp160/gp120 (53) were positive in our assay. The six patients who sero converted showed a low positivity by ELAVIA-1 (optical density near the cutoff value) reacted by blocking-ELAVIA-1 with an McAb binding inhibition greater than 85%. By contrast, negative samples (29) and specimens that exhibited reactivity only against gag-proteins (10) were not detected (McAb binding inhibition smaller than 15%). This sensitive and specific blocking-ELAVIA-1 represents a convenient alternative to WB as a confirmatory test. The technique is time-saving and inexpensive and can easily be integrated with a screening test for diagnostic or epidemiologic studies on HIV-1 infection.

A new C1q solid-phase assay for the detection of IgM immune complexes: application to the follow-up of kidney graft recipients with recurrent cytomegalovirus infection.

A C1q solid-phase enzyme assay for detecting circulating immune complexes (CIC) containing immunoglobulins M was developed. IgM immune complexes (IgM-CIC) bind to purified C1q-coated microplates. The amount of bound IgM-CIC is determined by adding anti-mu-alkaline phosphatase conjugate. This assay proved IgM-CIC specific; it does not detect IgG-CIC nor free anti-CMV IgM. Furthermore Rheumatoid Factor (RF) does not interfere with this test. Then, 119 renal allograft recipients were followed-up for the presence of seric IgM-CIC. Among them, 86 patients developed a recurrent Cytomegalovirus (CMV) infection and 33 did not. In the non-infected population, IgM-CIC were detected in only 15.2% of the patients. In contrast, patients with recurrent CMV infection showed a significantly higher frequency of detectable IgM-CIC (62.8%) (p less than 0.01). These IgM-CIC were detected: (a) during the first two or three weeks after graft; (b) in the course of the second month post graft. This second peak of IgM-CIC was never observed in subjects non-infected with CMV. IgM-CIC occurred before or at the same time as the detectable anti-CMV IgM and virus excretion in urines. Presence of IgM-CIC was not influenced by graft rejection episodes. Such a marker might help in discriminating immune response to viral infection from immune graft rejection.

Pattern of anti-cytomegalovirus IgM antibodies determined by immunoblotting. A study of kidney graft recipients developing a primary or recurrent CMV infection.

In order to improve the knowledge of the humoral immune response to CMV infection, we developed an immunoblotting technique which allowed a better analysis of the changes in the pattern of anti CMV-polypeptides IgM. We examined 234 sera belonging to 27 renal allograft recipients developing a primary or recurrent CMV infection and 12 non infected recipients. Thus we found that 11 main anti CMV-polypeptides IgM antibodies were present in over 25% of the infected patients. They reacted with proteins whose molecular weights ranged from 32K to 205K. We showed that anti-p 45-47 IgM antibodies were present in 100% of CMV infected recipients and never in the non-infected population. They appeared very early in the course of the infection (5.43 weeks post-graft for primary infection and 5.00 weeks for recurrent ones) and, therefore, constitute a good marker of active infection. Two other CMV-specific IgM antibodies (anti-p 60-64 and anti-p 100) were found exclusively in the course of primary infections. Anti-p 60-64 IgM was observed at a high frequency (57.1%) and with a mean delay of 6.57 weeks post-graft. Therefore, the anti-p 60-64 IgM detection could be helpful for the diagnosis of primary infection. In almost 100% of both primary and recurrent infections, we observed anti-p 140 and anti-p 38 IgM antibodies. Only about 50% of non-infected patients had low levels of anti-p 140 and anti-p 38 IgM. The follow-up of recurrent infections showed that the anti CMV-polypeptides IgM antibodies appeared earlier than in primary infection. When we compared anti-p 45-47 IgM detection by immunoblotting and anti-CMV IgM detected by ELISA we observed that immunoblotting permitted the diagnosis 2.5 weeks earlier for primary infection, and 1 week earlier for recurrent infection, than ELISA. In addition, the detection of anti-p 45-47 IgM antibodies also occurred earlier than virus excretion.

[Immunologic markers of the risk of cytomegalovirus infection among immunosuppressed patients]. / Marqueurs immunologiques du risque d'infection à cytomégalovirus chez les immunodéprimés.

ELISA and complement fixation test (CF) were compared for the determination of the Cytomegalovirus (CMV) humoral immune status of donors and recipients before and after graft. ELISA-IgG was more sensitive than the CF test and permitted a better identification of the high risk donors and recipients. The presence of IgM antibodies in the recipient at the day of transplantation is an index of greater susceptibility to chronic or recurrent infection at a later date. In primary renal allograft recipient the detection of specific IgM antibodies by ELISA provides more rapid diagnosis than that by CF. In bone marrow transplants, ELISA-IgM showed a better correlation with the virus isolation than the CF test. In the reinfected renal allograft patients the presence of IgM was related to clinical manifestations. Using ELISA, specific decreases of CMV antibody titers were detected before virus isolation.

[Immunological diagnosis of cytomegalovirus infections. Application to blood transfusion centers]. / Diagnostic immunologique des infections à cytomégalovirus. Application aux centres de transfusion sanguine.

The Blood Transfusion Centers (B.T.C.) are mainly concerned with the selection of CMV infection free blood donors, the screening of the anti CMV antibody high titre plasma donors and the evaluation of specific anti CMV Immunoglobulin preparations. Various serological methods could be used but they are of different value depending the purposes of the B.T.C. The neutralization test (Nt), with the addition of complement is specific and detects the protecting AB against the glycoproteins of the viral envelope. The complement fixation test (CF) using extracts of CMV infected cells as antigen largely varies in its sensitivity according to the quality of the antigen. In any case, the CF test is not sensitive enough to detect a latent CMV infection in a certain percentage of the non immunosuppressed adults, but could be used for the selection of anti CMV antibody high titres carriers. Three sensitive methods: passive haemagglutination, indirect immunofluorescence and indirect ELISA tests, might be used for the detection of latent CMV infections. They detect various AB against various internal and external components of the CMV. They are submitted to various sources of errors. The sensitivity of the indirect IF test is mainly restricted by the quality of the antigen preparation, its specificity by the presence of anti cells antibodies in the sera, the Fc receptors in the antigens and the specificity of the conjugates. The indirect ELISA which is submitted to the same causes of errors is a highly sensitive test, easy to perform, reagents are available, and automatic processors have been developed. When compared with the previous techniques, the ELISA test is suitable for the screening of CMV free donors, when it is performed with an highly sensitive antigen. It could be also used for the screening of high antibody titre carriers: its correlation with the CF test is quite good (r = 0,82). When comparatively applied to the titration of Immunoglobulins preparations, made from plasmas or placentas, for either IM or IV administration, the Elisa test gives AB titres different from those obtained with Nt and indirect IF. The calibration of a standard Immunoglobulin reagent is urgently needed and double blind clinical assays of the protective effect of various preparations of specific anti CMV Immunoglobulins have to be promptly designed.