CMV DNA is rarely detected in healthy blood donors using validated PCR assays

BACKGROUND: Although serologic screening or WBC reduction of blood components can reduce the incidence of transfusion-transmitted CMV (TT-CMV) infection, 'breakthrough' cases of TT-CMV still occur and may produce serious sequelae. NAT of blood components for CMV DNA has been proposed to further reduce the risks of TT-CMV. However, large-scale studies to determine the utility of validated CMV NAT assays for donor screening have not been reported. STUDY DESIGN AND METHODS: Coded whole-blood samples (n=1000) were tested for the presence of CMV DNA using two CMV PCR assays previously validated in a multicenter trial (a nested PCR assay directed at the CMV UL93 open-reading frame and the Roche Monitor assay). Corresponding plasma samples were tested in parallel for the presence of anti-CMV using other assays (Abbott CMV EIA and Fujirebio/Olympus CMV particle agglutination assays). RESULTS: In total 416 and 514 of the samples tested as CMV-seropositive and -seronegative, respectively, by both antibody assays. The remaining 70 samples had discrepant serology results. Only 2 of the 1000 samples (both seropositive) had reproducibly detectable CMV DNA (positive in at least three of four replicates). CMV DNA was not reproducibly detected in seronegative samples or in samples with discrepant serology results. CONCLUSIONS: Although previous investigations showed frequent detection of CMV DNA in healthy CMV-seropositive (and some seronegative) blood donors, these studies were relatively small and the performance characteristics of their assays were difficult to evaluate. In contrast, the present large cross-sectional study of US donors utilized two previously validated PCR assays and demonstrated that CMV DNA is only rarely detectable in seropositive donors. Thus, the use of CMV PCR assays with optimal performance characteristics did not increase the detection of potentially infectious blood components beyond that provided by current serologic screening assays.     

Cytomegalovirus DNA can be detected in peripheral blood mononuclear cells from all seropositive and most seronegative healthy blood donors over time

BACKGROUND: A poor correlation between cytomegalovirus (CMV) seroreactivity and the risk of CMV transmission prompted an investigation of the presence of CMV DNA in peripheral blood mononuclear cells (PBMNCs) from seropositive and seronegative blood donors. Because latent CMV exists in monocytes, monocyte-enriched cells were analyzed separately. STUDY DESIGN AND METHODS: Samples from 270 blood donors were tested with a sensitive polymerase chain reaction (PCR) test that detects two CMV genes, and the results were correlated to CMV serology. Cross-reactivity with other herpesvirus genes was not recorded. RESULTS: PCR testing demonstrated that 71 percent of seropositive donors harbor CMV in PBMNCs. Thus, not all seropositive donors were CMV DNA positive when individual samples were tested. Tests repeated over a period of time showed that all seropositive individuals were positive. Increased sensitivity was obtained with enriched monocytes. Among seronegative donors, 55 percent harbored CMV DNA in monocyte-enriched PBMNCs, while 14 percent had CMV DNA in PBMNCs. CONCLUSION: All seropositive donors harbored latently infected PBMNCs, as demonstrated by the testing of samples collected over time. In addition, a substantial proportion of seronegative individuals are CMV carriers and might transfer infection. The findings concur with clinical evidence of CMV transmission by blood components from seronegative individuals and with in vitro reactivation of CMV in PBMNCs from seronegative donors.     

Duration of time from onset of human immunodeficiency virus type 1 infectiousness to development of detectable antibody. The HIV Seroconversion Study Group

Background : For persons newly infected with the human immunodeficiency virus type 1 (HIV-1), the time from the onset of infectivity to the development of detectable HIV-1 antibody is unknown. Persons who donate blood during this period account for nearly all instances of HIV-1 transmission from HIV-1 antibody-screened blood transfusions. Study Design and Methods : To estimate the window period from infectivity to HIV-1 antibody positivity, 701 HIV-1-seropositive blood donors who made a previous seronegative donation at 40 United States blood centers were studied. The HIV-1 antibody status was determined for at least one recipient of blood from the seronegative donation preceding the seropositive donation made by 182 of the 701 donors. Results : There were 39 seropositive recipients of blood from these 182 donors. Three donors were excluded from further analysis because the seropositive recipients of their blood had other HIV-1 risk factors or had HIV-1 infection before transfusion. The final study population comprised the remaining 179 donors, of whom 36 (20%) transmitted HIV-1 infection to recipients. When the interval between the seropositive donation and the preceding seronegative donation was less than 180 days, 46 percent of the donors transmitted HIV-1. In contrast, when that interval exceeded 540 days, only 2 percent transmitted HIV-1. A mathematical model was developed to explain the relationship between the probability that the previous seronegative donation occurred during the donor's window period of infectiousness, and hence transmitted HIV-1, as a function of both the window period and the duration between the seropositive and previous seronegative donations. This model indicated that the transmission data were most consistent with an average window period of 45 days. Assuming a log-normal window period distribution, it was estimated with 95 percent certainty that at least 90 percent of persons had a window period of less than 141 days. Conclusion : The window period averages 45 days, with few, if any, donors remaining infectious and seronegative for longer than 6 months.     

Frequency and duration of plasma CMV viremia in seroconverting blood donors and recipients

BACKGROUND: Both CMV-seronegative blood and unscreened, filtered blood carry a low but definite risk of transmitting CMV infection. To explain this residual risk, evidence of cell-free viremia was sought in seroconverting and seroprevalent blood donors and seroconverting transfusion recipients by means of a plasma-based assay for CMV DNA. STUDY DESIGN AND METHODS: A CMV DNA PCR assay (COBAS Amplicor CMV Monitor, Roche) was used to detect CMV DNA in 384 paired plasma samples from 192 donors who seroconverted to anti-CMV, 488 anti-CMV EIA-positive samples from 60 seroprevalent donors, and 113 serial samples from 11 seroconverting recipients with posttransfusion CMV hepatitis. RESULTS: Three of 384 samples from 192 seroconverting donors had low levels of plasma CMV DNA (400-1600 copies/mL); one donor was positive before seroconversion, and the other two, after seroconversion. None of the 488 serial samples from 60 anti-CMV- positive donors contained CMV DNA in plasma. Three of 11 recipients demonstrated transient plasma viremia that temporally coincided with seroconversion. CONCLUSIONS: Plasma CMV DNA was detected in a small percentage of seroconverting blood donors and a larger percentage of recipients but was undetectable in seroprevalent donors. Plasma viremia in seroconverting donors may partially explain the low residual risk of CMV transmission by both CMV-seronegative and WBC-reduced seropositive blood.     

Recognition of cytomegalovirus clinical isolate antigens by sera from cytomegalovirus-negative blood donors

BACKGROUND: The most common way to prevent transmission of CMV by blood transfusion is to use blood products from seronegative donors. Screening of blood donors for CMV infection is usually based on detection of antigens obtained from the CMV laboratory strain AD 169. Recent evidence suggests that approximately up to 20 percent of CMV-negative blood donors may in fact be CMV-DNA positive by PCR analyses. STUDY DESIGN AND METHODS: In this study, sera from CMV-seronegative, CMV-seropositive, and CMV-DNA-positive/seronegative individuals, and from patients with acute and convalescent CMV infection for detection of CMV antibodies were analyzed. CMV antigens prepared from cells infected with CMV clinical isolates or the CMV laboratory strain AD 169 in ELISA and Western blot assays were used. RESULTS: All CMV-positive sera from blood donors were seropositive for the CMV antigens prepared from AD 169 (A2) or from a CMV clinical isolate (C6). Interestingly, whereas all CMV-negative blood donors were negative in tests for the CMV antigen A2, 36 percent were CMV seropositive using the CMV antigen C6 in ELISA. CONCLUSION: The data suggest that a substantial number of CMV-seronegative/CMV-DNA-positive serum samples contain antibodies that recognize CMV clinical isolate antigens.     

Multicenter evaluation of PCR methods for detecting CMV DNA in blood donors

BACKGROUND: CMV DNA screening may be a useful adjunct to serologic tests in distinguishing potentially infectious blood donations from those that are "CMV-safe." However, there is currently no consensus on the optimal assay method for accurate detection of CMV DNA in donors. STUDY DESIGN AND METHODS: A blinded multicenter evaluation of seven CMV PCR assays was performed by five laboratories by using coded sets of analytical controls and donor blood samples. RESULTS: Five assays displayed sufficient sensitivity for donor screening, as judged by consistent detection of a minimum of 25 CMV genome equivalents (geq) in analytical controls constructed to contain from 1 to 100 CMV geq in background DNA from 250,000 cells, while the other two assays displayed inadequate sensitivity. Three sensitive assays, two based on nested PCR directed at the UL93 and UL32 regions of the CMV genome and another test (Monitor Assay, Roche), did not detect CMV DNA in samples from any of 20 pedigreed CMV-seronegative, Western blot-negative (S-/WB-) donors. Two other assays based on nested PCR occasionally detected CMV DNA in S-WB- samples, and one sensitive nested PCR assay directed at UL123 detected CMV DNA in a large proportion (85%) of S-WB- samples. CONCLUSION: Seven CMV PCR assays currently used for research and/or diagnostic applications displayed marked variations in sensitivity, specificity, and reproducibility when applied to coded analytical and clinical control samples containing cellular DNA from the equivalent of 250,000 WBCs. These results will be useful in the selection of assays with performance characteristics appropriate to donor screening objectives. They may also help explain discrepant findings from previous studies that used PCR to determine CMV DNA prevalence in seronegative and seropositive blood donors.     

Validation of an automated immunoglobulin G-only cytomegalovirus (CMV) antibody screening assay and an assessment of the risk of transfusion transmitted CMV from seronegative blood

BACKGROUND: Cytomegalovirus (CMV) antibody donor screening assays have predominantly included both immunoglobulin G (IgG) and immunoglobulin M (IgM) detection. However, since in the majority of cases both CMV IgG and IgM are detected concomitantly during early seroconversion, CMV assays based only on IgG are now widely applied for donor screening.
STUDY DESIGN AND METHODS: The performance of an automated microparticle CMV IgG assay (Abbott AxSYM CMV IgG microparticle enzyme immunoassay [MEIA]) was compared with an established total antibody blood screening assay (Abbott CMV Total AB EIA). Sensitivity and specificity were assessed using 5050 random blood donors and 13 seroconversion panels. A risk analysis was undertaken to estimate the residual risk of transfusion-transmitted CMV (TT-CMV) from presumptive seronegative blood components.
RESULTS: The EIA achieved marginally (but not significantly) better resolved sensitivity (100%) than the AxSYM IgG assay (99.93%). The AxSYM IgG resolved specificity (99.34%) was superior to the EIA (96.4%). This superiority was maintained (98.61%) when a modified cutoff was applied to the AxSYM IgG assay to achieve 100 percent resolved sensitivity. The seroconversion sensitivities of the EIA and the AxSYM IgG were equivalent, detecting the same bleed as positive in the majority of the seroconversion panels tested. The median TT-CMV residual risk estimate for the two assays was approximately 1 in 66,000 (range, 42,000-165,000).
CONCLUSION: The AxSYM IgG MEIA is suitable for blood donor screening and was optimized by applying a modified cutoff of 9 AU per mL. The modeling predicts that implementing the AxSYM IgG assay would not negatively impact the already very low risk of TT-CMV associated with seronegative blood components in Australia.     

Transfusion- and community-acquired cytomegalovirus infection in children with malignant disease: a prospective study

BACKGROUND: The use of cytomegalovirus (CMV)-"safe" blood has been recommended for CMV seronegative patients with newly diagnosed malignant disease for whom bone marrow transplantation is a future option. STUDY DESIGN AND METHODS: To evaluate this policy, 76 CMV- seronegative children with lymphoreticular malignancies or solid tumors were randomly assigned to receive either blood components that were not screened for CMV antibody or CMV-seronegative red cell (RBC) and platelet units. Subjects were followed for evidence of CMV infection by the use of enzyme-linked immunosorbent assays and virus isolation. Follow-up continued long after the blood transfusions to determine the risk of community-acquired CMV infection. RESULTS: No cases of transfusion-acquired CMV infection were documented. The prevalence of CMV IgG and IgM antibody in blood donors was 40.5 and 0.9 percent, respectively. Patients assigned to receive standard blood components and CMV-negative components were given a median (range) of 7 (1-30) and 9 (1-38) RBC units and 11 (0-123) and 14 (0-71) platelet units, respectively. The risk of transfusion-acquired CMV infection is estimated to be less than 1 in 698 donor exposures. Two patients developed asymptomatic community-acquired CMV infection, for an incidence of 1.7 percent per patient-year of follow-up. CONCLUSION: The risk of transfusion-acquired CMV infection in this population is low, largely because of the patients' low level of exposure to seropositive blood and the use of relatively white cell-reduced components for purposes other than CMV prevention. Such children at this center therefore continue to receive standard blood components. Strategies to prevent CMV seroconversion in these children should include parental education to minimize the risk of community-acquired infection.     

Age-related prevalence of cytomegalovirus IgG antibody in north-west England

We assessed the prevalence of cytomegalovirus (CMV) IgG antibody in north-west England. The prevalence of antibody was low in pre-school children, but rose rapidly on school entry (age 5 years). About 40% of female blood donors of child-bearing age were seropositive. Hospitalised young adults had a higher prevalence of antibody than blood donors of similar age. Our results emphasised the need for local data on CMV seroprevalence when planning the counselling of women about acquisition of CMV during pregnancy and when planning the provision of CMV seronegative blood and blood products.     

Fluctuation of HCV viral load before seroconversion in a healthy volunteer blood donor

BACKGROUND: Newly implemented NAT has been shown to be able to effectively identify HCV-positive blood donated during the preseroconversion period. STUDY DESIGN AND METHODS: EDTA-plasma pools of 24 donations were tested using an HIV-1/HCV multiplex NAT under an FDA-approved IND application. Samples in a positive pool were retested individually. Positive samples were further tested by two discriminatory assays to determine specific viral reactivity. Upon obtaining informed consent, seronegative donors with positive NAT results were enrolled into a follow-up study for risk factor analysis and laboratory testing. RESULTS: A donation by a 29 year old female was identified as HCV NAT-positive with negative serology and an elevated ALT. Her two previous donations, 5 and 12 months earlier, were both seronegative and with normal ALT. Her husband tested positive for HCV RNA. The donor remained seronegative for at least 36 days. The index donation had a viral RNA concentration of >500,000 copies per mL while the first seropositive sample was NAT-negative. Laboratory data on serial follow-up samples showed 100- to 1,000-fold fluctuations in viral load during a period of 48 days prior to seroconversion. CONCLUSIONS: This case suggests that, at least in some newly infected individuals, the HCV viral load can fluctuate dramatically prior to seroconversion, and that NAT, even on individual samples, will not totally prevent HCV transmission.     

Prevention of transfusion-associated cytomegalovirus infection in very low-birthweight infants using frozen blood and donors seronegative for cytomegalovirus

During a 15-month period, 34 low birthweight infants (less than 1300 g) lacking maternal antibody to cytomegalovirus (CMV) received transfusions from an average of 10 donors per infant. Blood products consisted of predominantly washed deglycerolized frozen red cells from donors lacking antibody to CMV (an average of nine seronegative donors per infant). None of these infants acquired CMV infection while hospitalized. The absence of acquired CMV infections in these infants was in marked contrast (p less than 0.001) to the incidence of CMV infections in seronegative low-birthweight infants during the previous 21-month period when 28 percent acquired CMV (7 of 25 infants). The infants in this previous group received transfusions from an average of 11 donors per infant with an average of four seropositive donors per infant. These results confirm that hospital-acquired CMV infections can be prevented for very low-birthweight infants by donor selection and/or blood processing.     

Transfusion-acquired cytomegalovirus infection in neonates. A prospective study

The incidence of cytomegalovirus (CMV) infection was determined in 114 transfused neonates of any birthweight born to CMV antibody-negative mothers. In a second phase of this study, an additional 28 transfused infants weighing less than 1250 g, born to both CMV antibody-negative and antibody-positive mothers, were followed. All infants underwent weekly virus culture and monthly serology during hospitalization and at 6 to 12 weeks after their last transfusion. Only one of 126 (0.8%) seronegative infants and one of 16 (6.3%) seropositive infants developed CMV infection. If the assumption is made that the CMV-infected infant received only 1 unit of infectious blood, the risk of transfusion-acquired CMV infection to seronegative infants is 0.16 percent per cellular unit transfused or 0.37 percent per seropositive cellular unit transfused. Despite similarities in the prevalence of CMV antibody in the donor population, the age of blood products used, and the mean number of donor exposures, a significantly lower incidence of CMV infection was found in the seronegative transfused infants than that in two previously published studies (p less than 0.01, p less than 0.001). Because no mortality and very little morbidity could be attributed to transfusion-acquired CMV infection in the nurseries, the authors can see no justification for the provision of specialized blood components for the prevention of CMV infection in this patient population.     

Human immunodeficiency virus type 1-infected blood donors: epidemiologic, laboratory, and donation characteristics. The HIV Blood Donor Study Group

Transmission of human immunodeficiency virus type 1 (HIV-1) by homologous blood transfusion in the United States (US) is minimized by the deferral of potential donors who are at risk for HIV-1 infection and by the screening of all donations for HIV-1 antibody. HIV-1-seropositive donors at 20 blood centers were studied for information to be used in evaluating the safety of the US blood supply and making recommendations to increase that safety. From June 1988 to August 1989, 829 (0.04%) of 2,192,000 donors were found to be seropositive; 512 were interviewed. Of 388 seropositive men, 56 percent had had sex with men, 10 percent had used drugs intravenously, 8 percent had had sex with intravenous drug users, and 27 percent had no identified risk. Of 124 seropositive women, 58 percent had had sex with men at risk for HIV (81% of whom used drugs intravenously), 5 percent had used drugs intravenously, and 41 percent had no identified risk. Racial and ethnic minorities made up 68 percent of seropositive donors (black, 38%; Hispanic, 30%) and approximately 14 percent of all donors. The 157 persons with no identified risk had demographic characteristics and serologic test results for syphilis and hepatitis B that were more similar to those of HIV-1-seropositive donors with recognized risk than to those of seronegative donors. Three health care worker-blood donors (from an estimated 93,100 health care worker-donors) had infection that was probably acquired occupationally.(ABSTRACT TRUNCATED AT 250 WORDS)     

High prevalence in Central Africa of blood donors who are potentially infectious for human herpesvirus 8

BACKGROUND: In western countries, the transmission of human herpesvirus 8 (HHV-8) via blood transfusion has been recently postulated. In sub- Saharan African, the incidence of HHV-8-associated Kaposi's sarcoma and the seroprevalence for HHV-8 in autochthonous populations are high. STUDY DESIGN AND METHODS: The aim of this study was to estimate the prevalence of blood donations potentially infectious for HHV-8 in the general adult population of Central Africa. Forty-nine blood donors at the Centre de Transfusion Sanguine in Bangui, the capital of the Central African Republic, were selected. Forty-five inpatients of Broussais Hospital, Paris, France, who were known to be seronegative for HIV and hepatitis B and C viruses and who had not received heart or kidney transplants, were chosen as a European "control" group for comparison. HHV-8 DNA sequences were detected in peripheral blood mononuclear cells by nested polymerase chain reaction using primer sets located in the HHV-8 open reading frame 26. RESULTS: Eleven (22.5%; 95% CI: 12%-37%) of 49 blood donors were positive for HHV-8. Three (6%) were HIV-1 seropositive. Two (67%) of the 3 HIV-infected blood donors were also positive for HHV-8. All blood donors were apparently healthy; none was known to suffer from Kaposi's sarcoma. Only one (2.2%) control was carrying HHV-8 DNA on the peripheral blood mononuclear cells. The prevalence of HHV-8 was higher in blood donors from Bangui than in patients from Broussais Hospital. CONCLUSIONS: HHV-8 infection is highly prevalent in an apparently healthy adult population from Central Africa, which raises concerns about HHV-8 transmission through transfusion in this setting.     

Multicenter comparison of serologic assays and estimation of human herpesvirus 8 seroprevalence among US blood donors

BACKGROUND: As part of assessing the possibility of transfusion transmission of human herpesvirus 8 (HHV-8 or Kaposi's sarcoma-associated herpesvirus), HHV-8 seroprevalence was estimated among US blood donors, the performance of HHV-8 serologic tests was compared, and the presence of HHV-8 DNA was tested for in donated blood. STUDY DESIGN AND METHODS: Replicate panels of 1040 plasma specimens prepared from 1000 US blood donors (collected in 1994 and 1995) and 21 Kaposi's sarcoma patients were tested for antibodies to HHV-8 in six laboratories. HHV-8 PCR was performed on blood samples from 138 donors, including all 33 who tested seropositive in at least two laboratories and 22 who tested positive in at least one. RESULTS: The estimated HHV-8 seroprevalence among US blood donors was 3.5 percent (95% CI, 1.2%-9.8%) by a conditional dependence latent-class model, 3.0 percent (95% CI, 2.0%-4.6%) by a conditional independence latent-class model, and 3.3 percent (95% CI, 2.3%-4.6%) by use of a consensus-derived gold standard (specimens positive in two or more laboratories); the conditional dependence model best fit the data. In this model, laboratory specificities ranged from 96.6 to 100 percent. Sensitivities ranged widely, but with overlapping 95 percent CIs. HHV-8 DNA was detected in blood from none of 138 donors evaluated. CONCLUSIONS: Medical and behavioral screening does not eliminate HHV-8-seropositive persons from the US blood donor pool, but no viral DNA was found in donor blood. Further studies of much larger numbers of seropositive individuals will be required to more completely assess the rate of viremia and possibility of HHV-8 transfusion transmission. Current data do not indicate a need to screen US blood donors for HHV-8.     

Failure to detect human cytomegalovirus DNA in peripheral blood leukocytes of healthy blood donors by the polymerase chain reaction

The transmission of cytomegalovirus (CMV) by blood transfusion may have a major effect on certain immunocompromised patients. To protect susceptible blood recipients from infection, it is advisable to use blood components from CMV-seronegative donors. However, serologic tests are not capable of indicating which blood component actually harbors infectious virus and can transfer it to the recipient. Therefore, a sensitive method is needed for the detection of the virus itself. There have been three reports on the detection of CMV in healthy volunteer blood donors by the polymerase chain reaction (PCR). CMV DNA was found in all seropositive and most seronegative blood donors. However, many other authors have failed to confirm these data. A highly sensitive and specific PCR assay was developed for the detection of CMV DNA in peripheral blood leukocytes. With this protocol, blood samples from 116 volunteer blood donors were investigated. None of these samples proved to be positive for CMV DNA. In contrast, CMV DNA was detected in 10 of 10 renal transplant patients early in the course of active CMV infection. It can be concluded that the CMV genome copy number in the peripheral blood leukocytes of healthy individuals is beyond the detection limit of current PCR technology.     

Screening of blood donors for idiopathic CD4+ T-lymphocytopenia

BACKGROUND: The recent recognition of idiopathic CD4+ T-lymphocytopenia (ICL) had led to concern that an unknown immunodeficiency virus may be transmissible by transfusion. STUDY DESIGN AND METHODS: To evaluate the prevalence and significance of low CD4+ values among blood donors, CD4+ data on 2030 blood donors who were negative for antibody to human immunodeficiency virus type 1 (HIV-1) were compiled. Those with CD4+ values below ICL cutoffs (< 300 CD4+ T cells/microL, or < 20% CD4+ T cells) were recalled for follow-up investigations. Serial CD4+ data on 55 homosexual men who seroconverted during prospective follow-up and data on 139 anti-HIV-1-positive blood donors initially evaluated in 1986 were reviewed as well. RESULTS: Five seronegative donors (0.25%) had absolute CD4+ counts < 300 cells per microL and/or < 20 percent. On follow-up, all five donors had immunologic findings within normal ranges, lacked HIV risk factors, and tested negative for HIV types 1 and 2 and human T-lymphotropic virus type I and II infections by antibody and polymerase chain reaction assays. Four of five donors reported transient illness shortly after their low CD4+ count donations. The median interval from HIV-1 seroconversion to an initial CD4+ value below ICL CD4+ cutoffs was 63 months for infected homosexual men. Of 139 HIV-1-infected blood donors studied 1 to 2 years after seropositive donations, 34 (24%) had CD4+ counts < 300 cells per microL and/or < 20 percent. CONCLUSION: Low CD4+ counts are rare among anti- HIV-1-negative volunteer blood donors and are generally associated with transient illnesses. If any unknown virus progresses similarly to HIV- 1, CD4+ count donor screening would be a poor surrogate for its detection.     

A search for hepatitis C virus polymerase chain reaction-positive but seronegative subjects among blood donors with elevated alanine aminotransferase

BACKGROUND: Previous studies reported the existence of hepatitis C virus (HCV) polymerase chain reaction (PCR)-positive but seronegative sera. This is not surprising in the case of window-phase specimens, because PCR can detect HCV RNA many weeks before the appearance of antibody. To determine whether such sera can also be found in chronically infected subjects, a high-risk population of blood donors with elevated alanine aminotransferase was studied. STUDY DESIGN AND METHODS: Freshly frozen plasma from 301 donors with alanine aminotransferase > 100 IU per L was tested with PCR assays that were rigidly controlled for specificity and contamination, and with current and newer versions of assays for anti-HCV. Sera were classified as seropositive if positive in two screening assays and one supplemental assay or if positive in two screening assays and PCR. RESULTS: New versions of screening assays detected 100 percent of seropositive samples. A second-generation immunoblot assay detected 98 percent of seropositive sera, a second-generation recombinant immunoblot assay detected 96 percent, and an enzyme immunoassay for antibody to the envelope protein of HCV detected 98 percent. Fifty-one of 54 seropositive sera were PCR positive. None of the 247 seronegative samples was reproducibly positive on PCR. CONCLUSION: No PCR-positive but seronegative donors were found in this high-risk donor population. The possible benefit of PCR screening of blood donors can be determined only by large-scale comparative testing of donor populations and may be limited to the detection of window-phase infections.     

The impact of blood transfusion on the occurrence of pneumonitis in primary cytomegalovirus infection after liver transplantation

The influence of blood transfusion, and particularly the transfusion of blood from cytomegalovirus (CMV)-seropositive donors, on the incidence of primary CMV infection following liver transplantation was studied prospectively in 29 consecutive CMV-seronegative adult liver transplant recipients. Fourteen patients received a liver graft from a CMV- seropositive donor (Group A), whereas for 15 patients, the donor was CMV seronegative (Group B). Eleven patients (79%) in Group A but only two (13%) in Group B developed CMV infection (odds ratio, 23.8; 95% confidence interval, 3.5-169.4). In five Group A patients, primary CMV infection resulted in pneumonitis. There was no statistical difference in the total number of blood units and the number of units of CMV- seropositive blood given to patients who did or did not develop CMV infection in both Groups A and B (odds ratio for unit of CMV- seropositive blood transfused, 1.07; 95% confidence interval, 0.96- 1.19). However, Group A patients who developed CMV pneumonitis received a higher total number of blood units (median, 44) and of CMV- seropositive blood units (median, 18) than did patients who developed other CMV infections (asymptomatic CMV, CMV syndrome, or CMV hepatitis), who received a median of 9 total units of blood and 5 units of CMV-seropositive blood (p = 0.004). It is concluded that the total number of blood units and of CMV-seropositive blood units transfused does not have an effect on the incidence of primary CMV infection after liver transplantation, but it does have an impact on the severity of the infection in recipients of a CMV-seropositive allograft.