Evaluation of different assays for the detection of parvovirus B19 DNA in human plasma

Baylis, Sally A.; Shah, Nita R.; Minor, Philip D.

doi:10.1016/j.jviromet.2004.05.011

Cited by 69 publications

(80 citation statements)

References 15 publications

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“…The molecular biology and clinical significance of the newly recognized erythrovirus types are under active study (13,(15)(16)(17). In line with the difficulties in detecting these viruses (16,18,19), two previously undescribed human parvoviruses have been identified recently among symptomatic patients (20,21). Indeed, the evolution rate of these mammalian single-stranded DNA viruses has turned out to be exceptionally high, comparable with that of RNA viruses (22,23).…”

mentioning

confidence: 99%

Bioportfolio: Lifelong persistence of variant and prototypic erythrovirus DNA genomes in human tissue

Norja

Hokynar

Aaltonen³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

248

216

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Human erythrovirus is a minute, single-stranded DNA virus causing many diseases, including erythema infectiosum, arthropathy, and fetal death. After primary infection, the viral genomes persist in solid tissues. Besides the prototype, virus type 1, two major variants (virus types 2 and 3) have been identified recently, the clinical significance and epidemiology of which are mostly unknown. We examined 523 samples of skin, synovium, tonsil, or liver (birth year range, 1913-2000), and 1,640 sera, by qualitative and quantitative molecular assays for the DNA of human erythroviruses. Virus types 1 and 2 were found in 132 (25%) and 58 (11%) tissues, respectively. DNA of virus type 1 was found in all age groups, whereas that of type 2 was strictly confined to those subjects born before 1973 (P < 0.001). Correspondingly, the sera from the past two decades contained DNA of type 1 but not type 2 or 3. Our data suggest strongly that the newly identified human erythrovirus type 2 as well as the prototype 1 circulated in Northern and Central Europe in equal frequency, more than half a century ago, whereafter type 2 disappeared from circulation. Type 3 never attained wide occurrence in this area during the past >70 years. The erythrovirus DNA persistence in human tissues is lifelong and represents a source of information about our past, the Bioportfolio, which, at the individual level, provides a registry of one's infectious encounters, and at the population level, a database for epidemiological and phylogenetic analyses.epidemiology ͉ gene therapy ͉ parvovirus ͉ phylogeny ͉ single-stranded DNA

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mentioning

confidence: 99%

Bioportfolio: Lifelong persistence of variant and prototypic erythrovirus DNA genomes in human tissue

Norja

Hokynar

Aaltonen³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

248

216

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“…The 32 B19V DNA-positive and the 13 B19V DNA-negative samples were arranged along an estimated hypothetical curve according to the B19V DNA load (IU/ml in ordinate axis) and the type of marker(s) detected (time of antibody response in abscissa axis), resulting in a profile of the evolution of B19V infection [6,8,11,12,14,16,17]. …”

Section: Methodsmentioning

confidence: 99%

“…B19V infection evolution is characterized by a 5-day phase with high viremia (titer about 10 14 IU/ml B19V DNA) [6,7,8]. This viremia is neutralized with antibodies generated by the humoral immune system directed against two structural viral proteins VP1 and VP2 [9,10].…”

Section: Introductionmentioning

confidence: 99%

Characterization of Markers of the Progression of Human Parvovirus B19 Infection in Virus DNA-Positive Plasma Samples

Bonjoch¹,

Obispo²,

Alemany³

et al. 2015

Transfus Med Hemother

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Background: Accurate characterization of the infection stage in parvovirus B19(B19V)-positive plasma donations would help establish the donation deferral period to contribute to a safe fractionation pool of plasma. Methods: Viral DNA load of 74 B19V DNA-positive plasma samples from whole blood donations was determined by titration using nucleic acid testing. Markers of cellular (neopterin) and humoral (B19V-specific IgM and IgG) immune response were determined by ELISA in 32 B19V DNA-positive samples and in 13 B19V DNA-negative samples. The infection progression profile was estimated according to B19V DNA load and the presence of immune response markers. Results: B19V DNA load in the 74 samples was 10⁶-10¹³ IU/ml. The distribution of 14 out of 32 selected B19V DNA-positive samples plus 2 B19V DNA-negative samples with no immune response marker followed along an upward curve according to B19V DNA load. After the peak, the distribution of 18 immune marker-positive samples followed along a downward curve according to their B19V DNA load and was grouped as follows: neopterin (n = 4), neopterin+ IgM (n = 8), neopterin + IgM + IgG (n = 3), IgM + IgG (n = 2), IgM (n = 1). There were 11 B19V DNA-negative IgG-positive samples. Conclusion: This study of B19V-DNA load and levels of neopterin, IgM, and IgG allows for reliable characterization and distribution into the different stages of B19V infection.

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“…However, diagnostic kits also need to face problems due to target heterogeneity, and for B19V some reports and proficiency panel evaluations suggest that qPCR assays may not always obtain uniform analytical performances (7,(25)(26)(27). On the other hand, the in-house development of molecular assays will need a high degree of conformity to analytical guidelines and validation procedures to ensure reliable results.…”

Section: B19v Genetic Variabilitymentioning

confidence: 99%

Keeping Pace with Parvovirus B19 Genetic Variability: a Multiplex Genotype-Specific Quantitative PCR Assay

et al. 2013

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Three genotypes have been identified within the parvovirus B19 species (B19V), and such genetic diversity may have significant implications for the development of molecular detection assays. In the present study, B19V genetic variability has been examined on a subset of genomic sequences available in the NCBI nucleotide database, and a quantitative PCR (qPCR) assay able to detect, differentiate, and quantify all viral variants has been established. The designed primers and probes have been used for the development of alternative detection formats, based on a combined use of intercalating dye and genotype-specific hydrolysis probes. The qPCR assay analytical performances have been determined on the 1st WHO International Reference Panel for Parvovirus B19 Genotypes. The developed qPCR protocols allow for the detection of genotypes 1 to 3 with equal accuracy, and with a limit of detection (LOD) of 200 IU/ml. A comparison of routine performance was carried out with respect to a previously established assay specifically validated on B19V genotype 1. For 130 clinical samples analyzed, 126 showed concordant results (31 positive and 97 negative), while 4 showed discordant results. Overall, the genotype-specific qPCR assay showed a sensitivity of 93.94% and a specificity of 97.94%, with an agreement rate of 96.92%. The proposed qPCR assay and the alternative protocols developed, each with robust performance, may allow choice with respect to operational systems and diagnostic requirements and might contribute to provide a more reliable diagnostic service and epidemiological surveillance of B19 virus. H uman parvovirus B19 (B19V) is a widespread human-pathogenic virus and a member of the Erythrovirus genus in theParvoviridae family (1). In the last decade, due to the introduction of innovative molecular techniques, the taxonomy in the family has grown in complexity to accommodate a still-increasing number of newly recognized viruses (2). Specifically, B19 viruses are now formally subdivided into three genotypes, the prototype genotype 1 and the two variant genotypes 2 and 3, and further subtypes (3, 4). In particular, within genotype 1, the majority of isolates are referred to as genotype 1a, while a few isolates from Asia show separate clustering and are referred to as genotype 1b (5). Within genotype 3, two distinct subtypes are usually referred to as genotypes 3a and 3b (6). All B19V genotypes appear to cocirculate, but their relative frequencies are strikingly different: genotype 1a is ubiquitous worldwide, genotype 2 is rare and sporadically detected in different geographic settings, including Europe, and genotypes 3a and 3b are detected at higher frequencies in western Africa and at lower frequencies in other geographic areas (7-10).The three B19V genotypes are assumed to have similar biological properties, pathogenic capacities, and transmission routes and pose a similar diagnostic challenge in the clinical setting (11). Of relevance, all genotype variants can be contaminants of blood and blood-derived products, so ...

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Evaluation of different assays for the detection of parvovirus B19 DNA in human plasma

Cited by 69 publications

References 15 publications

Bioportfolio: Lifelong persistence of variant and prototypic erythrovirus DNA genomes in human tissue

Bioportfolio: Lifelong persistence of variant and prototypic erythrovirus DNA genomes in human tissue

Characterization of Markers of the Progression of Human Parvovirus B19 Infection in Virus DNA-Positive Plasma Samples

Keeping Pace with Parvovirus B19 Genetic Variability: a Multiplex Genotype-Specific Quantitative PCR Assay

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