Dec 22, 2015
Are plasma derivatives Are plasma derivatives sources for parvovirus B19 sources for parvovirus B19
transmission?transmission?
Manal M. BaddourManal M. BaddourAssociate Professor of Microbiology Associate Professor of Microbiology
King Saud UniversityKing Saud University
Heegaard and Brown, 2002
Parvovirus B19 is an erythrovirus and Parvovirus B19 is an erythrovirus and was until recently the only known was until recently the only known member of the member of the Parvoviridae Parvoviridae family of family of viruses causing disease in humans. viruses causing disease in humans.
D e n so v irus
Ite ra v irus
B re v id e n so v irus
P e fun d en sov irus
D e n sov irin ae(S u b fa m ily)
M V M
C a n ine p a rvo v irusC P V
P o rc ine p a rvo v irusP P V
Parvovirus
A A V s-1 -?
G o ose pa rvov irusG P V
A v ia n A A VA A A V
Dependovirus
P a rvov irus B 19B 1 9V
S im ian p a rvo v irusS P V
P m P V
Erythrovirus
B o v in e pa rvov irusB P V
M inute v iru s of canine sM V C
H u m an b o ca v irus
Bocavirus
A le u tio n d ise a se(m in ks )
P a rv4 /P a rv5
Am dovirus
Parvovirinae(S u b fa m ily)
Parvoviridae(F a m ily )
Parvovirus B19 (B19) was discovered Parvovirus B19 (B19) was discovered fortuitously in 1973 in the United Kingdom fortuitously in 1973 in the United Kingdom by Cossart, who was looking for the by Cossart, who was looking for the hepatitis B virus in blood from “healthy” hepatitis B virus in blood from “healthy”
donors.donors. Cossart et al, 1975Cossart et al, 1975
The virus selectively targets early erythroid The virus selectively targets early erythroid precursors, causing anemia. precursors, causing anemia.
The cellular receptor is the blood group P The cellular receptor is the blood group P antigen antigen
Brown et al., 1993Brown et al., 1993
After infection, the incubation period ranges from After infection, the incubation period ranges from 6 to 18 days. 6 to 18 days.
Viremia in early infection occurs with extremely Viremia in early infection occurs with extremely high titers (up to 10high titers (up to 101111 to 10 to 101313 genome genome equivalents/ml). equivalents/ml).
In immunocompetent persons, viraemia is In immunocompetent persons, viraemia is usually cleared a few weeks after infection. usually cleared a few weeks after infection.
In immunocompromised patients, the virus can In immunocompromised patients, the virus can cause a persistent infection for several months cause a persistent infection for several months and years mainly due to impairment of B cell and years mainly due to impairment of B cell function.function.
Siegl and Cassinotti, 1998Siegl and Cassinotti, 1998
The frequency of B19-DNA blood donors has The frequency of B19-DNA blood donors has been estimated at rates of 1:167 to 1:35,000 been estimated at rates of 1:167 to 1:35,000
Heegaard & Heegaard &
Brown,2002Brown,2002
High seroconversion rates and some High seroconversion rates and some cases of symptomatic illness have been cases of symptomatic illness have been due to blood products (eg immune due to blood products (eg immune globulin, clotting factors) prepared from globulin, clotting factors) prepared from B19-containing plasma pools with reports B19-containing plasma pools with reports of prominent B19 infection and worsening of prominent B19 infection and worsening the patient’s condition as a resultthe patient’s condition as a result
Hayakawa et al, 2002; Yee et al, 1995;Hayakawa et al, 2002; Yee et al, 1995;
Zanella et al 1995; Mortimer et al, 1983Zanella et al 1995; Mortimer et al, 1983
Aim of This StudyAim of This Study
To determine the frequency of parvovirus To determine the frequency of parvovirus B19 DNA contamination in a range of B19 DNA contamination in a range of plasma derivatives commercially available plasma derivatives commercially available in the local market for clinical use.in the local market for clinical use.
MaterialMaterial
Samples from three batches of each of the Samples from three batches of each of the following types of products from 2 companies following types of products from 2 companies were tested for parvovirus DNA by nested PCR:were tested for parvovirus DNA by nested PCR:
1- human albumin1- human albumin 2- human coagulation factor VIII2- human coagulation factor VIII 3- human coagulation factor IX (one company)3- human coagulation factor IX (one company) 4- human normal IV immunoglobulins 4- human normal IV immunoglobulins Serum from a patient previously diagnosed to be Serum from a patient previously diagnosed to be
positive for parvovirus B19 DNA by PCR was positive for parvovirus B19 DNA by PCR was used as a control.used as a control.
MethodsMethods DNA from each of these products was DNA from each of these products was
extracted in triplicates from 200µl aliquots extracted in triplicates from 200µl aliquots of the tested products using the of the tested products using the QIAamp QIAamp DNA Blood MinikitDNA Blood Minikit
Nested PCR using primers Nested PCR using primers • A1 5´-AATGAAAACTTTCCATTTAATGA-3´ A1 5´-AATGAAAACTTTCCATTTAATGA-3´ • B1 5´-TCCTGAACTGGTCCCGGGGATGG-3´ B1 5´-TCCTGAACTGGTCCCGGGGATGG-3´ 590 bp590 bp• A2 5´-GCGTGGAAGTGTAGCTGTGCC-3´ A2 5´-GCGTGGAAGTGTAGCTGTGCC-3´ • B2 5´-CAGAGCTTTCACCACCACTGC-3´B2 5´-CAGAGCTTTCACCACCACTGC-3´355 bp 355 bp
Frickhofen and Young, 1989
Carriere et al, 1993
A representative sample of each type A representative sample of each type of product was spiked with parvovirus of product was spiked with parvovirus B19 DNA (extracted from serum of a B19 DNA (extracted from serum of a previously determined B19 positive previously determined B19 positive patient) and subjected to the same patient) and subjected to the same procedures to test for the efficiency of procedures to test for the efficiency of the technique.the technique.
Results, Results, cont.cont.
Donor Plasma Donation Center
Begin Manufacturing
Complete Manufacturing
Patient
Safety Step 9:Post-Marketing
Surveillance
Safety Step 1:Donor
Screening
Safety Step 2:Testing Donation
Safety Step 3:Inventory Hold and Lookback
Safety Step 4:Plasma Pool
Testing
Safety Step 7:Virus Inactivation
Virus/TSE Removal
Safety Step 5 & 6:Quality
AssuranceGMP
Safety Step 8:GCP
Packaging Guidance
Manufacturing SafeguardsManufacturing SafeguardsPathogen Safety is a comprehensive approach with effective redundant measures that provide a high margin of safety
The Plasma Protein Therapeutics Association
Pre-implementation
Implementation of Parvovirus B19 NAT Testing
PROCESSVIRAL REDUCTION
PROCESSVIRAL REDUCTION
No Testing UnknownMargin ofSafety
IncreasedMargin ofSafety
Reduction in Parvovirus B19 Viral LoadsReduction in Parvovirus B19 Viral Loads
101- 109
IU/mL
Production Pool
<105
IU/mL
Production PoolMini-pool Testing96-1200 donation samples
High Titer Plasma
ConclusionsConclusions It is true that results obtained from the present It is true that results obtained from the present
study seem reassuring, probably due to study seem reassuring, probably due to implementation of strict screening of blood implementation of strict screening of blood donors or plasma pools for parvovirus B19 DNA donors or plasma pools for parvovirus B19 DNA by the plasma products' manufacturing by the plasma products' manufacturing companies. companies.
Nevertheless, continuous monitoring of blood Nevertheless, continuous monitoring of blood products should be a routine procedure to products should be a routine procedure to ensure safety of such products for recipients ensure safety of such products for recipients especially those at risk to avoid the hazardous especially those at risk to avoid the hazardous possibilities of serious infections.possibilities of serious infections.
Conclusions, Conclusions, cont.cont.
Human bocavirus HBoVHuman bocavirus HBoV B 19 genotypes 2 (A6) & 3 (V9)B 19 genotypes 2 (A6) & 3 (V9)