Infecção congênita é aquela transmitida da mãe ao feto antes do nascimento. A transmissão vertical pode ocorrer por via transplacentária ou por contato direto com o patógeno durante o parto. A fonte de infecção é o microrganismo presente no sangue da gestante durante a infecção primária ou crônica. Estima-se que as infecções perinatais representam 2% a 3% de todas anomalias congênitas e as mais comuns são representadas pela sigla TORCH, que inclui Toxoplasmose, Outras (como sífilis e varicela-zoster), Rubeóla, Citomegalovírus e Herpes. A maioria das infecções TORCH causa morbidade materna leve, assintomática, mas tem consequências fetais graves e, em geral, o tratamento da infecção materna não tem impacto sobre o resultado fetal. Por isso, o reconhecimento do contágio materno e o monitoramento fetal são de extrema importância na prevenção de doenças congênitas. O presente projeto visa o desenvolvimento de um produto multiparamétrico utilizando a tecnologia xMAP®, criada pela companhia norte-americana Luminex Corporation, para detecção simultânea de anticorpos da classe IgG anti-Toxoplasmose, anti-Rubéola e anti- Citomegalovírus em amostras de sangue coletado em papel de filtro. O produto, inédito no mercado nacional, tem o objetivo de atender à demanda específica da triagem pré-natal no país. Como objetivos específicos pode-se citar: a melhoria da eficiência dos programas de triagem pré-natal, graças à economia de tempo, amostras e reagentes; a contribuição financeira para o Brasil, uma vez que será produzido nacionalmente, gerando emprego e renda; a possibilidade de ampliar o mercado a partir do desenvolvimento futuro de novos produtos baseados na mesma metodologia. Para tanto, utilizaram-se antígenos específicos acoplados à microesferas de poliestireno e anticorpos de detecção conjugados à estreptavidina-ficoeritrina. Foram analisadas 1499 amostras de gestantes, coletadas em papel de filtro, cedidas e previamente triadas pela APAE-Goiânia, com o objetivo de comparar os resultados obtidos através da análise com o protótipo desenvolvido com aqueles já confirmados pelo laboratório utilizando a tradicional metodologia de ELISA. Os resultados de Concordância e Sensibilidade foram superiores a 78% para todos os parâmetros. Por outro lado, os valores de Especificidade foram mais baixos, principalmente para os parâmetros Rubéola e Citomegalovírus. Importante ressaltar o pequeno número de amostras com resultado negativo para Citomegalovírus e Rubéola disponível, o que refletiu diretamente no cálculo da Especificidade do produto / Congenital infections are transmitted from the mother to the fetus before birth. Vertical transmission can occur via placenta or by direct contact with the pathogen during childbirth. The source of infection is the microorganism present in the pregnant woman\'s blood during primary or chronic infection. It is estimated that perinatal infections are responsible for 2% to 3% of all congenital abnormalities and the most common are represented by the acronym TORCH, including Toxoplasmosis, Others (such as syphilis and varicella-zoster), Rubella, Cytomegalovirus (CMV) and Herpes. Most infections TORCH causes mild maternal morbidity, asymptomatic, but has serious fetal consequences fetal and, generally, maternal infection treatment has no impact on fetal outcome. Therefore, the recognition of maternal infection and fetal monitoring are extremely important in preventing birth defects. This project aims to develop a product using the multiplex xMAP® technology, created by US company Luminex Corporation, for simultaneous detection of IgG antibodies anti-toxoplasmosis, antirubella and anti-cytomegalovirus in blood samples collected in filter paper. The product, unprecedented in brazilian market, aims to meet the specific demand of prenatal screening in Brazil. The specific objectives are: improving the efficiency of prenatal screening programs, thanks to savings in time, samples and reagents; the financial contribution for Brazil, as it will be produced nationally, generating jobs and income; the possibility of expanding the market from the future development of new products based on the same methodology. For this purpose, were used specific antigens coupled to polystyrene beads and antibodies conjugated to streptavidin-phycoerythrin. Were analyzed 1499 samples of pregnant women, collected on filter paper, pre-screened by APAE-Goiânia, in order to compare the results obtained from the analysis with the prototype developed with those already confirmed by the laboratory using traditional ELISA methodology. The results of Concordance and Sensitivity were higher than 78% for all parameters. In contrast, the Specificity values were lower, especially for Rubella and Cytomegalovirus parameters. Importantly the small number of negative samples negative for Cytomegalovirus and Rubella provided by APAE-Goiânia, which is directly reflected in the product specificity value.

Les péri-implantites se caractérisent par une destruction inflammatoire irréversible des tissus qui entourent l’implant dentaire. Leur pathogénie est proche des parodontites, maladies inflammatoires chroniques d’étiologie bactérienne entrainant une destruction irréversible de l’os alvéolaire soutenant les dents. Cependant, la réponse inflammatoire et la résorption osseuse diffèrent entre péri-implantites et parodontites. De plus, les traitements conventionnels non chirurgicaux ne sont pas efficaces pour le traitement des péri-implantites. L’objectif de notre travail est d’analyser les différences entre les médiateurs inflammatoires et cataboliques des tissus péri-implantaires en comparaison aux tissus parodontaux malades après une thérapie conventionnelle non chirurgicale. Nous avons bâti une collection biologique d’explants humains de tissus conjonctifs péri-implantaires ou parodontaux malades. Une analyse Multiplex des milieux conditionnés et histologiques des explants a révélé une expression accrue de l’inhibiteur de métalloprotéase TIMP-2, de l’interleukine IL-10 et du RANK-L dans les tissus péri-implantaires. En accord avec une revue de littérature que nous avons mené dans la seconde partie de notre travail, notre hypothèse est que ces différences s’expliqueraient, tout du moins en partie, par la présence de particules de titane au niveau des tissus péri-implantaires. Notre travail participe à la compréhension de la pathogenèse de la maladie et concourt à l’identification de biomarqueurs potentiels pour le pronostic ou le développement d’approches thérapeutiques. / Peri-implantitis lesions present an irreversible destruction of the peri-implant surrounding tissues. The pathogenic pathway of peri-implantitis is close from the one of periodontitis which is a chronic inflammatory disease caused by a bacterial biofilm leading to irreversible supportive bone destruction around teeth. Still, the inflammatory process and the bone destruction differ between the two diseases. In addition, conventional non-surgical treatments are not effective for the treatment of peri-implantitis. The aim of our work is to analyze the differences between inflammatory and catabolic mediators of peri-implant tissues compared to diseased periodontal tissues after conventional non-surgical therapy. We have constructed a biological collection of human explants of peri-implant or periodontal connective tissue. Multiplex assays of the conditioned media and histological analysis of the explants revealed an increased expression of the TIMP-2 metalloprotease inhibitor, interleukin IL-10, and RANK-L in the peri-implant tissues. In agreement with a review of literature that we conducted in our work, our hypothesis is that these differences can be explained, at least partly, by the presence of titanium wear particles in the peri-implant tissues. Our work contributes to the understanding of the pathogenesis of the disease and contributes to the identification of biomarkers.

This thesis discusses the technique of flow cytometry for multiplex analysis and its use in conjunction with imunochemical methods. As part of this work was carried out clinical studies dealing with secondary prevention of myocardial infarction and atherosclerosis in 186 pacientů. In this time represents myocardial infarction worldwide civilizational problem. A number of possible parameters for monitoring atherosclerosis in the world is still an unresolved issue. In the practical part of this work we performed an analysis using Luminex xMAP technology for new parameters (adiponectin, resistin, osteopontin) to predict atherosclerotic disease associated with myocardial infarcion. Also we wanted to see how these parameters are changed in patients after increasing the dose of therapeutic drugs.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Ying Fang / The Porcine Disease Complex (PDC) results in major economic problems for swine producers. PDC outbreaks result in increased mortality, decreased feed efficiency, higher cull rates, prolonged days to market and increased treatment costs. This disease involves the interaction and participation of many multifactorial etiologies including both bacterial and viral organisms playing a role in disease initiation and progression. The most common viral pathogens associated with the PDC include porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus (PCV2) and swine influenza virus (swIV). The recent outbreak of porcine epidemic diarrhea virus (PEDV) in the US swine herd has made the PDC even more complicated. In aid of the prevention and control of the PDC, veterinarians and producers require fast and efficient diagnostic tests for controlling the disease. In this study, we have generated recombinant nucleocapsid antigens to these viruses for use in a Luminex™ technology-based fluorescent microsphere immunoassay (FMIA). Utilizing these recombinant nucleocapsid antigens, the FMIA was developed to simultaneously detect antibodies in serum from animals infected with PEDV, PRRSV, SwIV and PCV2. The FMIA was developed based on testing experimentally derived standard positive and negative control sera, and the diagnostic specificity and sensitivity were compared to that generated from the classical enzyme-linked immunosorbent assay (ELISA) or hemagglutination inhibition (HI) test. Based on an evaluation of 4147 serum samples with known serostatus, the multiplex FMIAs reached greater than 97.5% sensitivity and 92.3 % specificity. Results showed that multiplexing did not affect the diagnostic sensitivity or specificity of each individual assay. This work provides a platform for the development of multiplex assays for detecting various swine pathogens simultaneously and aids in preventing and controlling the PDC.

Identification of the complement C4d fragment in peritubular capillaries as a specific marker for antibody mediated rejection in renal transplantation revealed the critical role of antibodies in graft survival. In this thesis, I document the design and findings of studies performed to investigate the clinical impact of anti-HLA antibodies present before and/or after transplantation. Over time, the detection techniques for anti-HLA antibodies has evolved from the less sensitive complement-dependent lymphocytotoxicity (CDC) crossmatching (XM) to more sensitive solid phase assays such as Luminex®. Studies have been conducted to compare the predictive value of different antibody detection techniques. The first result chapter presents antibody specificity in positive CDC B-cell crossmatch (BXM), analysed with highly specific Luminex® assays. The study also investigates the predictive value of BXM in the general transplant population. I found that donor-specific anti-HLA antibodies (DSA) are only present in one third of positive BXM and are associated with poor outcomes. The novel finding is that >80% of the DSA detected by BXM are complement-fixing IgG₁ and IgG₃ subclasses. Transplant glomerulopathy (TG) is type of chronic renal graft rejection. The pathogenesis of TG is unclear. In the second result chapter, I report risk factors and involvement of anti-HLA antibodies in the development of TG. This study shows that glomerular rejection, delayed graft function, HLA presensitization and DSA have a univariate effect on TG development. Multivariate analysis revealed that DSA are an independent predictor of TG, after adjustment for other risk factors. I have further investigated the role of BXM in a unique, well-matched, highly sensitized patient group transplanted under the national renal exchange programme. I compared Luminex® antibody analysis with BXM in predicting transplant outcomes. In highly sensitized patients, DSA are found in two thirds of positive BXM. In univariate analyses, BXM is associated with humoral rejection whereas DSA defined by Luminex® are associated with total and all rejection types. The major finding is that, by multivariate analysis, DSA defined by Luminex® are an independent predictor of total and humoral rejection, but BXM are not. These interesting findings are reported in the third result chapter. Studies reported in this thesis define the clinical significance of anti-HLA antibodies in renal transplant outcomes. Method comparison studies provide useful information on antibody specificity and their impact on graft survival. Collectively, a better understanding of alloantibodies associated with graft rejection and limitation of antibody detection methods may facilitate donor selection and choice of immunosuppressants, and consequently improve transplant outcomes. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1379925 / Thesis (Ph.D.) - University of Adelaide, School of Medicine, 2010

La majorité des modèles d’uvéites expérimentales sont médiées par les sous-types lymphocytaires CD4+ T-Helper 1 (Th1) et Th17. Le but de cette thèse est d’étudier les mécanismes immunitaires mis en jeu lors d’une uvéite idiopathique (sans étiologie) chez l’humain. La première partie de cette étude est donc consacrée à l’étude du profil biologique de 30 cytokines/chimiokines/ facteurs de croissance intraoculaires (AqH) et sériques obtenues au laboratoire (technologie Luminex®). Ces médiateurs sont dosés dans le sérum et l’AqH de patients atteints d’uvéites intermédiaires et postérieures activesidiopathiques. Ces résultats sont comparés à une cohorte de patients non inflammatoires et à des groupes de patients atteints d’uvéites de diverses étiologies. Nous avons trouvé que le profil des médiateurs diffère nettement en fonction de la cause de l’inflammation intraoculaire. Une réponse pro-inflammatoire large est observée avec des concentrations accrues des cytokines de type Th1 (IFNγ, IL-12), Th17 (IL-17), TNF-α, IL-6, MIP-1α, MIP-1β, MCP-1, G-CSF et IP-10 dans les uvéites idiopathiques. Les concentrations sériques différent de celles des contrôles par les taux d’IL-17 et IP-10 augmentés. La deuxième partie concerne l’étude des mécanismes immunitaires des uvéites par l’analyse des vascularites en imagerie adaptative (FIOA) et particulièrement l’observation anatomique des modifications liées aux inflammations oculaires. Nous ne trouvons pas de corrélation entre l’aspect de cellularité périvasculaire en FIOA et les cytokines retrouvées dans l’AqH des yeux avec uvéite. Pouvoir déterminer quels médiateurs de l’inflammation interviennent dans les pathologies inflammatoires permettrait de les cibler avec des thérapies adaptées. / Most of experimental models of uveitis are mediated by lymphocytic subtypes CD4+ T-Helper 1 (Th1) et Th17. The aim of this thesis is to study the immune mechanisms in idiopathic uveitis (i.e. unknown etiology) in humans. First, we will determine the biological spectrum of 30 cytokines/ chemokines/ growth factors in aqueous humour and sera by Luminex® technology. Those mediators will be measured in the serum and aqueous humour of patients with active intermediate and posterior uveitis classified as idiopathic. Those results will be compared to a cohort of patients with no inflammation and to patients with uveitis of various causes. We found that the profile of mediators changes regarding to the cause of the intraocular inflammation. The proinflammatory process is related to increased levels of cytokines Th1 (IFNγ, IL-12), Th17 (IL-17), TNF-α, IL-6, MIP-1α, MIP-1β, MCP-1, G-CSF and IP-10 idiopathic uveitis. The sera of the patients with idiopathic uveitis have increased levels IL-17 and IP-10. Secondly, the immune mechanisms of uveitis were studied by adaptive optics imaging (FIOA) and particularly the observation of anatomic changes related to ocular inflammations. We ruled out any correlation betweeen the perivascular cellular opacification in FIOA and the cytokines/ chemokines found in the eyes with uveitis. An understanding of the profile of mediators in inflammation and their determination could help to target idiopathic uveitis with correct therapies.

Today, a lot of proteomic research is aimed at discovering disease specific proteins. This requires theavailability of high-throughput, ultra-sensitive protein detection methods. Compartmentalized immunosequencing(cI-Seq) is a proximity-independent immuno-polymerase chain reaction (IPCR) based proteindetection method. Antigen recognition in cI-Seq is mediated by antibody pairs in which one of theantibodies is conjugated to a DNA-probe. The affinity recognition events occur in emulsion droplets inwhich the DNA-probes will be amplified through emulsion PCR (emPCR) and thereafter analyzed usingMassively Parallel Sequencing (MPS). The amplifiable nature of the DNA-probes improves the sensitivityof the detection, while the use of emulsion droplets and MPS increases the multiplex capacity andthroughput. Ultimately, cI-Seq enables analysis and detection even of lowly abundant proteins therebyincreasing the probability of discovering novel disease specific proteins. In this project, conjugation of DNA probes to antibodies was performed through two different approaches;Covalent Conjugation and Conjugation using Biotin and NeutrAvidin. Both of these approaches showedadvantageous and disadvantageous features. However, neither of them succeeded in producing stableconjugates in an efficient and reproducible manner. After conjugation, the DNA-conjugated antibodieswere used in immune complex formation. However, the immune complexes either failed to form or wereformed in an inefficient manner.

No description available.

La pathogenèse de la sacculite anale demeure incomprise. Cette condition semble cependant être plus fréquente chez les chiens atopiques. La sacculite anale se traite principalement avec un antibiotique. Avec la montée de l’antibiorésistance, il est important de mieux comprendre sa pathogenèse afin de prévenir la maladie et trouver des traitements alternatifs aux antibiotiques. Cette étude visait donc à comparer le microbiote bactérien et les cytokines pro-inflammatoires dans les sacs anaux de chiens sains à ceux de chiens atopiques, afin de déterminer si des changements pourraient être à l’origine des sacculites anales chez les chiens atopiques. L’hypothèse de ce projet était que le microbiote bactérien et les cytokines pro-inflammatoires dans les sacs anaux des chiens sains diffèrent de ceux des chiens atopiques traités ou non traités. Les principaux objectifs de cette étude étaient donc de caractériser le microbiote bactérien (MB) des sacs anaux des chiens atopiques recevant ou non un traitement (oclacitinib, cetirizine ou immunothérapie allergénique) et de déterminer si la concentration de certaines cytokines pro-inflammatoires libérées dans les sacs anaux variait entre les chiens sains et les chiens atopiques non traités (CANT) et les chiens atopiques traités (CAT). Des écouvillons floqués ont été utilisés pour échantillonner le rectum et les sécrétions provenant des sacs anaux de 15 chiens sains, six CAT et 14 CANT pour l’analyse du microbiote bactérien. L’extraction de l’acide désoxyribonucléique a été effectuée avec le kit commercial DNeasy PowerSoil. Le séquençage de la région V4 du gène de l’acide ribonucléique ribosomique 16S a ensuite été réalisé avec la plateforme Illumina MiSeq. Pour l’analyse des cytokines, le contenu des sacs anaux de 15 chiens sains, 12 CANT et cinq CAT a été prélevé dans un microtube. Chaque microtube a été mélangé au vortex. Le surnageant a ensuite été prélevé. Les microtubes contenant le surnageant ont ensuite été congelés à -80°C jusqu’à leur analyse. La concentration de l’interféron gamma, des interleukines (IL)-2, 6, 8, 10 et 12/23p40, de la protéine chimiotactique 1 des monocytes, du facteur de croissance des nerfs bêta, du facteur de cellules souches, du facteur de nécrose tumorale alpha (TNF-α) et du facteur de croissance de l’endothélium vasculaire A a été mesurée avec le test multiplex de Luminex. L’appartenance à la communauté était différente entre les sacs anaux des chiens sains et des CANT, des chiens sains et des CAT et des CANT et des CAT (P = 0,002, P = 0,013 et P = 0,012, respectivement). La structure de la communauté était différente entre les chiens sains et les CANT (P = 0,003) et entre les CANT et les CAT (P = 0,017), mais pas entre les chiens sains et les CAT (P = 0,332). Toutes les cytokines pro-inflammatoires évaluées ont été détectées dans les sacs anaux de chiens sains, de CANT et de CAT. Il n’y avait aucune différence significative entre les groupes, excepté pour l’IL-8 et le TNF-α, où l’IL-8 était plus élevée dans les sacs anaux des CANT en comparaison avec ceux des CAT (P = 0,02), et le TNF-α était en concentration plus faible dans les sacs anaux des chiens sains comparativement aux sacs anaux des CAT (P = 0,04). Il y a une dysbiose dans les sacs anaux de chiens atopiques, ce qui pourrait expliquer pourquoi les chiens atopiques semblent prédisposés à développer des sacculites anales bactériennes. Les traitements (oclacitinib, cetirizine et immunothérapie allergénique) semblent également modifier la composition du microbiote bactérien dans les sacs anaux des chiens atopiques pour qu’elle se rapproche de celle des chiens sains. L'IL-8 pourrait également jouer un rôle dans le développement de la sacculite anale. D’autres études évaluant un plus large nombre de cytokines permettraient possiblement de mettre en évidence des cytokines ayant un rôle important lors de sacculite anale chez les chiens atopiques. / The pathogenesis of anal sacculitis is not well understood. However, this condition appears to be more common in atopic dogs. Anal sacculitis is primarily treated with antibiotic therapies. With the rise of antimicrobial resistance, it is important to better understand the pathogenesis of anal sacculitis in order to prevent the disease and find alternative treatments to antibiotics. The present study aimed to compare the bacterial microbiota and pro-inflammatory cytokines in the anal sacs of healthy dogs with those of atopic dogs, in order to determine if there are changes that could explain anal sacculitis in atopic dogs. The hypothesis of this project was that the bacterial microbiota and proinflammatory cytokines in the anal sacs of healthy dogs differ from those of treated and untreated atopic dogs. The main objectives of this study were therefore to characterize the bacterial microbiota of the anal sacs of atopic dogs receiving or not a treatment (oclacitinib, cetirizine or allergen-specific immunotherapy) and to determine if the concentration of certain proinflammatory cytokines released in the anal sacs differed between healthy, untreated atopic dogs (UAD) and treated atopic dogs (TAD). Flocked swabs were used to sample the rectum and secretions from the anal sacs of 15 healthy dogs, six TAD and 14 UAD for bacterial microbiota analysis. Deoxyribonucleic acid extraction was performed with the commercial DNeasy PowerSoil kit. Sequencing of the V4 region of the 16S ribosomal ribonucleic acid gene was then performed with the Illumina MiSeq platform. For cytokine analysis, the contents of the anal sacs of 15 healthy dogs, 12 UAD, and five TAD were collected in a microtube. Each microtube was vortexed before the supernatant was collected. The microtubes containing the supernatant were then frozen at -80°C until analysis. The concentration of interferon gamma, interleukin (IL)-2, 6, 8, 10, and 12/23p40, monocyte chemotactic protein 1, nerve growth factor beta, stem cell factor, tumor necrosis factor-alpha (TNF-α), and vascular endothelial growth factor-A were measured with the Luminex multiplex assay. Community membership was different between the anal sacs of healthy dogs and UAD, healthy dogs and TAD, and UAD and TAD (P = 0.002, P = 0.013, and P = 0.012, respectively). Community structure was different between healthy dogs and UAD (P = 0.003) and between UAD and TAD (P = 0.017), but not between healthy dogs and TAD (P = 0.332). All proinflammatory cytokines assessed were detected in the anal sacs of healthy dogs, UAD, and TAD. There were no significant differences between groups except for IL-8 and TNF-α. IL-8 was higher in UAD anal sacs compared to TAD anal sacs (P = 0.02) and TNF-α was in lower concentration in healthy dog anal sacs compared to TAD anal sacs (P = 0.04). There is a dysbiosis between the anal sacs of UAD and TAD which might explain why atopic dogs seem predisposed to bacterial anal sacculitis. Treatments received by atopic dogs (oclacitinib, cetirizine and allergen-specific immunotherapy) shift the microbiota of the anal sacs towards that of healthy dogs. IL-8 may also play a role in the development of anal sacculitis. Further studies on a larger number of cytokines may identify cytokines that are important in the pathogenesis of anal sacculitis in atopic dogs.

Foodborne illnesses are a significant public health challenge in the United States, with an estimated 9.4 million illnesses annually attributed to the consumption of contaminated food, of which 59% are estimated to be caused by viruses, 39% by bacteria and 2% by parasites. Timely detection and identification of the pathogens causing foodborne outbreaks is vital for the implementation of outbreak control strategies, allowing public health officials to prevent additional illnesses and maintain confidence in the food supply. Public health laboratories employ a variety of traditional and molecular testing techniques to identify foodborne outbreak etiologic agents. One technology is the Luminex XMap® microsphere system, which is also marketed as the Bio-Plex™ 200. This platform has a multiplexing capability with the potential to simultaneously detect up to 100 targets in one reaction. The studies described here show that the combination of two Bio-Plex assays with real-time virus assays and one extraction method provides a flexible foodborne outbreak screening algorithm that potentially identifies an outbreak-associated pathogen on the first day of specimen submission and aids in focusing confirmatory laboratory testing. In these studies, two microsphere-based assays were designed for use on the Bio-Plex 200 system as screening assays for the detection of four enteric protozoa (Giardia intestinalis, Cyclospora cayetanensis, Cryptosporidium parvum, Entamoeba histolytica) and six virulence determinants of shiga toxin-producing Escherichia coli (STEC), enterotoxigenic Escherichia coli (ETEC), enteroinvasive Escherichia coli (EIEC) and Shigella spp. Precision and limits of detections were established for both assays. The sensitivity and specificity of the protozoan assay as compared to reference methods ranged from 81.25% to 100% for most targets, while sensitivity for the E. histolytica target was 42.86%. Sensitivity and specificity for the bacterial assay was 100% as compared to reference methods. However, cross-reactivity of the protozoan assay E. histolytica target with E. dispar and of the bacterial assay uidA target with enteropathogenic E. coli strains was noted. Additionally, real-time detection of norovirus and rotavirus nucleic acids extracted with the QIAamp DNA Stool Mini Kit was statistically comparable to detection when extracted with the Ambion® MagMAX™-96 Viral RNA Isolation Kit combined with the KingFisher® Magnetic Particle Processor.