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Transfusion-transmitted disease.

Many patients with haemophilia are infected with viruses, due to treatment with blood products--particularly from large pool clotting factor concentrates before 1985. AIDS in haemophilic patients was first described in 1982 and it has significantly reduced the life expectancy of these patients. Although no new sero-conversions have occurred since 1986, management of HIV in haemophilia remains a clinical challenge. Transfusion-associated hepatitis was recognized in 1943, and it is now an important complication of haemophilia treatment. Vaccination against HAV is recommended. Intensively-treated older haemophilic patients usually have serological evidence of HBV infection. HBV transmission has been stopped, but hepatitis B vaccination is still practised, because HDV requires HBV for propagation. Many patients are infected with HCV: before 1985 almost all patients who received clotting factor concentrate developed non-A, non-B hepatitis, now recognized as HCV. Treatment strategies are being developed for HCV in haemophilic patients. Parvo virus can be transmitted by clotting factor concentrate; it is very resistant to sterilization processes, transmission causing severe illness even in immuno-competent individuals. New blood-borne viruses responsible for sero-negative hepatitis include: GBV-A, B and C, and HGV. Although there is no link between CJD and haemophilia, there is concern about possible blood product transmission.

Blood↗

[Viral risks associated with blood transfusion].

The presence of viruses in blood cells or plasma from asymptomatic donors is the major risk of transmitting an infectious agent through blood transfusion. The main viruses involved are hepatitis viruses and retroviruses. The risk of transmitting hepatitis B virus (HBV) and hepatitis C virus (HCV) has been progressively and efficiently reduced in the last years by the successive introduction of hepatitis B surface antigen (HBsAg) screening, elevated serum alanine aminotransferase (ALT), antibody to hepatitis B core (HBc Ab), and more recently antibody to hepatitis C virus (HCV Ab). The risk of transmitting human retroviruses like human immunodeficiency virus (HIV) and human T-cell leukemia/lymphoma virus (HTLV) has also been reduced drastically thanks to screening for corresponding antibodies. However, except for HBs Ag screening, the immunoassays used for HCV, HIV, or HTLV only detect antibodies. Therefore, although they are infectious, a few blood units may be not discarded. The efficacy of preventive measures depends on the incubation time, the infectivity during this silent phase, and the sensitivity of screening procedures. Human cytomegalovirus (HCMV) and parvovirus B 19 are responsible for common infections. Consequently, 30% to more than 50% adults have serologic evidence of past infection. However, both viruses may cause severe primo-infections in some circumstances, especially in pregnant women or immunodeficient individuals.

Cytomegalovirus Infections↗

Quantification of parvovirus B19 DNA using COBAS AmpliPrep automated sample preparation and LightCycler real-time PCR.

The COBAS AmpliPrep instrument (Roche Diagnostics GmbH, D-68305 Mannheim, Germany) automates the entire sample preparation process of nucleic acid isolation from serum or plasma for polymerase chain reaction analysis. We report the analytical performance of the LightCycler Parvovirus B19 Quantification Kit (Roche Diagnostics) using nucleic acids isolated with the COBAS AmpliPrep instrument. Nucleic acids were extracted using the Total Nucleic Acid Isolation Kit (Roche Diagnostics) and amplified with the LightCycler Parvovirus B19 Quantification Kit. The kit combination processes 72 samples per 8-hour shift. The lower detection limit is 234 IU/ml at a 95% hit-rate, linear range approximately 10(4)-10(10) IU/ml, and overall precision 16 to 40%. Relative sensitivity and specificity in routine samples from pregnant women are 100% and 93%, respectively. Identification of a persistent parvovirus B19-infected individual by the polymerase chain reaction among 51 anti-parvovirus B19 IgM-negative samples underlines the importance of additional nucleic acid testing in pregnancy and its superiority to serology in identifying the risk of parvovirus B19 transmission via blood or blood products. Combination of the Total Nucleic Acid Isolation Kit on the COBAS AmpliPrep instrument with the LightCycler Parvovirus B19 Quantification Kit provides a reliable and time-saving tool for sensitive and accurate detection of parvovirus B19 DNA.

Antibodies, Viral↗

Human parvovirus B19 infection within a family and risk for pregnant women.

During an outbreak of parvovirus B19 infection among four related families at least 70% of the household contacts, including a woman at the 33rd week of pregnancy, became infected. Twins were born at the 39th week of pregnancy, both with B19 infection. B19 DNA was detected in their sera by a nested PCR, anti-B19 IgM was detectable only by an immunofluorescence assay, and low levels of maternal anti-B19 IgG were demonstrable by an immunoenzymatic test in the serum of both children. All the haematological parameters were normal at birth and 6 months later, when B19 DNA and anti-B19 antibody were no longer detectable in serum samples. This observation emphasizes the high risk of B19 infection among household contacts and the possibility of a favourable outcome of the foetal infection, possibly related to infection late in the pregnancy.

Adult↗

Parvovirus B19 outbreak on an adult ward.

In November and December 1992, an outbreak of parvovirus B19 infection occurred among patients and staff on an adult mixed surgical ward at a large hospital in London. Three patients and 15 staff members were serologically confirmed as acute cases. The attack rate among susceptible members of staff was 47%. In those infected, arthralgia (80%) and rash (67%) were the most common symptoms. Of six susceptible in-patients on the ward, three became infected. One of the in-patients who had carcinoma of the mouth was viraemic for more than 10 days with marrow suppression resulting in the postponement of chemotherapy until intravenous immunoglobulin was given and he was no longer viraemic. Control measures taken included closure of the ward to new admissions, transfer of only immune staff to the ward, and restriction of the ward nursing staff to working only on that ward. Although no specific exposure was conclusively identified as a risk factor, there was a suggestion of an increased risk of acquiring parvovirus B19 infection among those staff who did not adopt strict hand washing procedures after each physical contact with a patient (RR = 2.33; P = 0.07). Knowledge of parvovirus B19 among interviewed health care workers was poor: only 42% reported knowing about parvovirus B19 and only 38% could name a patient category at risk of a severe outcome following infection. This is the first report of a nosocomial outbreak affecting an adult ward and of possible transmission of parvovirus B19 infection from staff to in-patients. Hospital control of infection teams should include parvovirus B19 in their outbreak containment plans.

Adult↗

A school outbreak of parvovirus B19 infection investigated using salivary antibody assays.

An outbreak of parvovirus B19 infection at a primary school was investigated using saliva samples. Antibody capture immunoassays for salivary B19 IgG and IgM were developed using a recombinant B19 antigen and monoclonal antibody to B19 virus. Evaluation of the salivary IgG assay using paired serum and saliva samples from 43 staff at St Thomas' Hospital showed that it had a sensitivity of 100% and a specificity of 95%. Evaluation of the salivary B19 IgM assay using 87 paired blood and saliva samples from a study of general practitioner rubella notifications showed it had a sensitivity of 60% and a specificity of 98%. Using the salivary assay the level of B19 IgG within 2 weeks of the start of the outbreak ranged from 5-33% in children and 29% in staff. By detecting salivary B19 IgM and/or B19 IgG seroconversions, attack rates of 8-50% in children in different classes and 47% in staff were observed. Household transmission was also studied and an attack rate of 45% was recorded in 11 susceptibles. After the outbreak, the level of B19 IgG in children with the highest attack rates was 60-70%, similar to that seen in adults in the UK. This study highlights the risk of B19 infection in an institutional setting and shows that saliva samples are a useful alternative to blood.

Adolescent↗

Low prevalence of antibody to human parvovirus B19 in Singapore.

A seroepidemiological survey was conducted in Singapore to assess the prevalence of antibody to human parvovirus B19. Sera were collected from 600 healthy individuals between 6 months and over 50 years of age and tested for IgG antibody against B19 virus by antigen capture indirect immunosorbent assay. The overall seropositivity rate was 16.2%. All the children under 5 years of age were seronegative. Antibody prevalence increased gradually from 3.5% in school children (5-14 years of age) to 7.7% in age group 15-19 years and then to 10.3% in young adults (20-24 years of age). In the age group 25-34 years the prevalence was 28% and in the age group over 35 years 65% had parvovirus B19 antibody. The results of the survey indicate that there has been very low incidence of B19 virus infection in Singapore during the last two decades.

Adolescent↗

Parvovirus B19 infection in pediatric solid-organ and bone marrow transplantation.

The clinical significance of parvovirus B19 infection in pediatric solid-organ and bone marrow transplanted patients is unclear. The overall prevalence of parvovirus B19 infection in these patients is about 1-2% during the first year after transplantation. The most common symptom is anemia, but leukopenia and thrombocytopenia have also been observed. Rare cases of hepatic dysfunction, myocarditis, vasculitis and respiratory failure have also been reported. Whereas serology is of limited value around the time of transplantation, it is recommended that a search for B19 DNA is included in first-line investigations in any transplanted patient with unexplained anemia. Specific antiviral therapy is not available, however, intravenous immunoglobulin produces rapid improvement in most cases. Although relatively rare, the severe complications following parvovirus B19 infection in the transplant setting can be avoided by early diagnosis and treatment.

Antibodies, Viral↗

Oral manifestations of papular-purpuric 'gloves and socks' syndrome due to parvovirus B19 infection: the first case presented in Greece and review of the literature.

Papular-purpuric 'gloves and socks' syndrome (PPGSS) is a novel, rare, self-limited dermatosis initially described in 1990. It is characterized by painful, pruritic edema and erythema, rapidly evolving to papular-purpuric lesions on the distal extremities, in a gloves-and-socks distribution, accompanied by fever and oral lesions such as petechiae, vesiculopustules and small erosions. Parvovirus B19 has been implicated in most cases as the etiological factor. Herein we present the first case of PPGSS in a 42-year-old Greek man with von Willebrand disease. On admission the patient was febrile, and presented acral edema and erythema rapidly followed by purpuric lesions on the same sites, and palatal petechiae. Complete remission of the exanthem occurred 7 days after hospitalization. Clinical and laboratory evaluation including serologic tests and PCR, confirmed the presence of parvovirus B19. Review of the existing literature on this novel syndrome and its association with parvovirus B19 is also presented.

Adult↗

Transmission of symptomatic parvovirus B19 infection by clotting factor concentrate.

The risk of acquiring diseases from transfusion of blood and blood products is well recognized and the issue of parvovirus in haemophiliacs is not a new one. We report two patients with haemophilia acquiring iatrogenic parvovirus B19 infection, resulting in life-threatening sepsis in one, and immunocompetent adult. Over the last 10 years there has been great progress in manufacturing safer products with regard to enveloped viruses such as HIV, hepatitis B and C. A recent outbreak across Europe of hepatitis A in haemophiliacs treated with plasma-derived factor VII concentrates has made haemophilic treaters concerned about the known (parvovirus B19 and hepatitis A) and the unknown non-lipid enveloped viruses that may be contained in the clotting factor concentrates, because these are resistant to the existing viral inactivating techniques. The possibility of HIV itself mutating into a non-lipid enveloped virus emphasizes the need to seek and use safer products.

Adult↗

Heat sensitivity of human parvovirus B19.

BACKGROUND AND OBJECTIVES: To date there has been no published report on a systematic evaluation of the heat sensitivity of human parvovirus B19 (B19) and the related safety of the plasma-derived fractionated products. In this study, we examined the heat sensitivity of B19 by using the infectivity assay with cultured cells. MATERIALS AND METHODS: The heat sensitivity of B19 was examined by measuring the reduction in viral infectivity titres after heating liquid containing B19 at 60 degrees C. Viral infectivity was assayed by detection of viral antigens or viral mRNA in infected cells. As a control, canine parvovirus (CPV) was also heat-treated. RESULTS: B19 displayed quite different inactivation kinetics to CPV when both were heated in liquid at 60 degrees C. In sharp contrast to the latter, B19 was rapidly inactivated within 1 h when the virus was suspended in 5% or 25% human serum albumin solution, phosphate-buffered saline, or complete medium. However, B19 appeared to be resistant to heat inactivation in liquid containing 60% sucrose. CONCLUSIONS: The heat sensitivity of B19 in liquid was clearly different from that of CPV. Significantly, the efficiency to inactivate B19 and reduce its infectivity following heating in liquid was mainly affected by the composition of the solutions used for virus suspension.

Antigens, Viral↗

Prevalence of human erythrovirus B19 DNA in healthy Belgian blood donors and correlation with specific antibodies against structural and non-structural viral proteins.

BACKGROUND AND OBJECTIVES: Human parvovirus (erythrovirus) B19 is recognized as a major contaminant of blood and blood products. To reduce the risk of contamination, plasma-pool screening and exclusion of highly viraemic donations are recommended. The objectives of this study were to estimate the prevalence of B19 DNA in our blood-donor population, to determine the appropriate pool size to be tested (taking into account parameters such as prevalence, viral load, test sensitivity, and the efficacy of inactivation procedures), and to correlate viral loads with the serological status of donors as regards antibodies against different viral proteins. MATERIALS AND METHODS: Pools of different sizes were tested for B19, using a sensitive nested polymerase chain reaction (PCR) as well as an simple, un-nested, less sensitive PCR. Positive pools were resolved to the level of individual donations, and the viral load and serological markers were determined. RESULTS: Of 16,859 donations, 27 (one of 625) were found to be B19 DNA positive, with viral loads ranging from 10(2) to > 10(7) IU/ml. Twenty-five of the positive donations were tested for VP-specific anti-B19 antibodies, and eight (32%) were negative for both immunoglobulin (Ig)M and IgG. They were probably collected in the preseroconversion window period or from chronic carriers without detectable antibodies. We regarded the seven (28%) IgM-positive donors as being in the early phase of infection. The remaining 10 (40%) IgM-negative, IgG-positive donors were probably carriers of persistent infection (i.e. PCR positive despite the presence of IgG antibodies), as suggested by their low viral loads (< 10(4) IU/ml). Fifteen out of 36 major pools contained one or more contaminated donations. Among these, 12 tested positive by nested PCR and only three by un-nested PCR, this reflecting a viral load of > 10(4) IU/ml. CONCLUSIONS: By testing all donations as pools of 480 by un-nested PCR, and resolving positive pools to identify the responsible donations, it is possible to ensure that the viral load in fractionation pools (5000 donations) remains < 10(3) IU/ml, compatible with the efficacy of inactivation procedures and complying with Food and Drug Administration (FDA) recommendations.

Algorithms↗