Existe uma crescente demanda por alternativas de controle de doenças para se reduzir o uso do controle químico, como por exemplo, o estudo da indução de resistência em plantas, através de fitotoxinas como a taxtomina A e outras substâncias. O vírus do mosaico do pepino (Cucumber mosaic virus - CMV) e o vírus do mosaico amarelo da abobrinha (Zucchini yellow mosaic virus - ZYMV) têm causado grandes prejuízos em culturas importantes, assim como o fungo Rhizopus stolonifer, que pode atacar diversos hospedeiros. Este trabalho foi conduzido com o objetivo de se avaliar os efeitos do uso da taxtomina A como indutor de resistência em plantas. Como resultado preliminar a produção de taxtomina A pode ser aumentada utilizando-se como fonte de inóculo amostra de uma cultura liquida pré-estabelecida de Streptomyces scabies. Foram testados a aspersão de taxtomina A em plantas de abobrinha de moita, para se avaliar a proteção contras os vírus CMV e ZYMV. Também foi aspergida taxtomina A em bagas de uva \"Itália\" para se avaliar o desenvolvimento da podridão pós-colheita causada por R. stolonifer. As plantas de abobrinha de moita, mantidas em casa de vegetação, tratadas com taxtomina A não apresentaram sintomas de mosaico característico do CMV. O teste ELISA apresentou resultados negativos indicando a não presença de partículas virais em níveis detectáveis nas plantas. Com relação ao ZYMV, a taxtomina A não foi capaz de controlar o desenvolvimento da doença, pois apareceram sintomas em todas as plantas. A taxtomina A apresentou efeito in vitro sobre R. stolonifer retardando o crescimento do fungo principalmente nas duas maiores concentrações usadas. Porém, a aplicação de taxtomina A não foi capaz de proteger as bagas de uva contra o patógeno. Finalizando a taxtomina A foi capaz de controlar em 100% a incidência de CMV em abobrinha, porém não foi capaz de controlar o ZYMV em abobrinha e a podridão nas uvas causada por R. stolonifer. / There is an increasing demand for alternative control of diseases to reduce the use of chemical control, for example the induced resistance studies on plants with phytotoxins like thaxtomin A and other substances. The Cucumber mosaic virus (CMV) and the Zucchini yellow mosaic virus (ZYMV) cause a lot of losses on important crops and also the fungus Rhizopus stolonifer that can cause disease in different hosts. This work was carried out to evaluate the use of thaxtomin A to induce resistance. Thus, as a preliminary result, the thaxtomin A production could be increased by using as inoculum an established Streptomyces scabies liquid culture. It was tested the spraying of thaxtomin A on zucchini plants to evaluate the protection against the viruses CMV e ZYMV. On the other hand, grape berries cv \"Italia\" were also sprayed with thaxtomin A to evaluate the development of the postharvest rot caused by R. stolonifer. The zucchini plants were maintained in the greenhouse and the treatment with thaxtomin A inhibited mosaic symptom development by the CMV. The ELISA test showed negative results indicating that there was no detectable virus particles inside the plants. The thaxtomin A was not able to control the ZYMV development since all plants exhibited symptoms. The thaxtomin A showed in vitro control of R. stolonifer as it reduced the fungal growing at the two highest concentrations used. However, the thaxtomin A was not able to control the postharvest rot in grapes. Finally, the thaxtomin A was able of controlling 100% of the incidence of CMV on zucchini plants, but was not able of controlling ZYMV in zucchini and the grape berrie rot caused by R. stolonifer.

Existe uma crescente demanda por alternativas de controle de doenças para se reduzir o uso do controle químico, como por exemplo, o estudo da indução de resistência em plantas, através de fitotoxinas como a taxtomina A e outras substâncias. O vírus do mosaico do pepino (Cucumber mosaic virus - CMV) e o vírus do mosaico amarelo da abobrinha (Zucchini yellow mosaic virus - ZYMV) têm causado grandes prejuízos em culturas importantes, assim como o fungo Rhizopus stolonifer, que pode atacar diversos hospedeiros. Este trabalho foi conduzido com o objetivo de se avaliar os efeitos do uso da taxtomina A como indutor de resistência em plantas. Como resultado preliminar a produção de taxtomina A pode ser aumentada utilizando-se como fonte de inóculo amostra de uma cultura liquida pré-estabelecida de Streptomyces scabies. Foram testados a aspersão de taxtomina A em plantas de abobrinha de moita, para se avaliar a proteção contras os vírus CMV e ZYMV. Também foi aspergida taxtomina A em bagas de uva \"Itália\" para se avaliar o desenvolvimento da podridão pós-colheita causada por R. stolonifer. As plantas de abobrinha de moita, mantidas em casa de vegetação, tratadas com taxtomina A não apresentaram sintomas de mosaico característico do CMV. O teste ELISA apresentou resultados negativos indicando a não presença de partículas virais em níveis detectáveis nas plantas. Com relação ao ZYMV, a taxtomina A não foi capaz de controlar o desenvolvimento da doença, pois apareceram sintomas em todas as plantas. A taxtomina A apresentou efeito in vitro sobre R. stolonifer retardando o crescimento do fungo principalmente nas duas maiores concentrações usadas. Porém, a aplicação de taxtomina A não foi capaz de proteger as bagas de uva contra o patógeno. Finalizando a taxtomina A foi capaz de controlar em 100% a incidência de CMV em abobrinha, porém não foi capaz de controlar o ZYMV em abobrinha e a podridão nas uvas causada por R. stolonifer. / There is an increasing demand for alternative control of diseases to reduce the use of chemical control, for example the induced resistance studies on plants with phytotoxins like thaxtomin A and other substances. The Cucumber mosaic virus (CMV) and the Zucchini yellow mosaic virus (ZYMV) cause a lot of losses on important crops and also the fungus Rhizopus stolonifer that can cause disease in different hosts. This work was carried out to evaluate the use of thaxtomin A to induce resistance. Thus, as a preliminary result, the thaxtomin A production could be increased by using as inoculum an established Streptomyces scabies liquid culture. It was tested the spraying of thaxtomin A on zucchini plants to evaluate the protection against the viruses CMV e ZYMV. On the other hand, grape berries cv \"Italia\" were also sprayed with thaxtomin A to evaluate the development of the postharvest rot caused by R. stolonifer. The zucchini plants were maintained in the greenhouse and the treatment with thaxtomin A inhibited mosaic symptom development by the CMV. The ELISA test showed negative results indicating that there was no detectable virus particles inside the plants. The thaxtomin A was not able to control the ZYMV development since all plants exhibited symptoms. The thaxtomin A showed in vitro control of R. stolonifer as it reduced the fungal growing at the two highest concentrations used. However, the thaxtomin A was not able to control the postharvest rot in grapes. Finally, the thaxtomin A was able of controlling 100% of the incidence of CMV on zucchini plants, but was not able of controlling ZYMV in zucchini and the grape berrie rot caused by R. stolonifer.

Nach einer Transplantation eines soliden Organs entwickeln 80 % aller Patienten eine Infektion. Hierbei nimmt die Infektion mit dem humanen Cytomegalievirus den größten Stellenwert ein. Verschiedene Studien weisen auf das Risiko hCMV-assoziierter Nierentransplantatabstoßungen hin. Ziel der vorliegenden Arbeit war es, das Transplantat- und Patientenüberleben der am Universitätsklinikum Leipzig nierentransplantierten Patienten unter Berücksichtigung der antiviralen sowie immunsuppressiven Medikation zur Verhinderung einer hCMV-Infektion auszuwerten. Die Ergebnisse wurden zur Entwicklung standardisierter Handlungsempfehlungen zur hCMV-Prophylaxe für Patienten nach einer Nierentransplantation genutzt. Im Zeitraum von 1993 - 2014 wurden 635 Patienten am Universitätsklinikum Leipzig nierentransplantiert. Insgesamt konnte bei 127 Patienten nach stattgehabter Nierentransplantation, bei einem medianen Nachbeobachtungszeitraum von 95 Monaten, eine hCMV-Infektion nachgewiesen werden. Darunter traten die meisten Infektionen in der Hochrisikokonstellation mit hCMV-negativen Empfängern auf, die ein hCMV-positives Nierentransplant erhielten. Dementsprechend ist der größte Risikofaktor für das Ausbilden einer hCMV-Infektion die Nierentransplantat-Spender- Empfänger-Konstellation. Zusätzlich kann als Risikofaktor ein hohes Lebensalter der Patienten und erhöhte Werte des Serumkreatinins zum Zeitpunkt der Entlassung aufgezeigt werden. Im Gegensatz zu den in der Literatur führenden Aussagen, kann an dem hier vorliegenden Patientenkollektiv die Induktionstherapie und die Art der Organspende nicht als Risikofaktor für eine hCMV-Infektion bestätigt werden. Es ist kein signifikanter Einfluss der hCMV-Infektion auf das Transplantatüberleben in der Untersuchung am gesamten Patientenkollektiv zu ermitteln. In der Untergliederung in die hCMV-Spender-Empfänger-Risikokonstellationen stellt sich jedoch in den Risikokonstellationen mit hCMV-negativen Donoren [(D-/R+), (D-/R-)] ein signifikant negativer Zusammenhang der hCMV-Infektion auf das Transplantatüberleben dar. In Bezug auf das Patientenüberleben kann sowohl für das gesamte Patientenkollektiv als auch in den hCMV-Spender-Empfänger-Risikokonstellationen mit hCMV-positiven Empfängern [(D+/R+), (D-/R+)] der signifikant negative Einfluss der hCMV-Infektion bestätigt werden. Zur Prävention einer hCMV-Infektion ist derzeit das oral zu verabreichende Medikament Valganciclovir das Mittel der Wahl. In der Hochrisikokonstellation (D+/R-) stellt sich ein Zusammenhang zwischen der Dauer der hCMV-Prophylaxe mit Valganciclovir und dem hCMV-freien Überleben dar. Dabei zeigt sich kein Vorteil einer 200-tägigen Valganciclovir-Prophylaxe im Vergleich zu einer 100-tägigen Prophylaxe. Je länger Valganciclovir eingesetzt wurde, umso höhere hCMV-Viruslasten traten zu einem späteren Zeitpunkt auf. Das Transplantatüberleben kann in der Hochrisikokonstellation (D+/R-) durch eine mindestens 100-tägige Valganciclovir-Prophylaxe signifikant verlängert werden im Vergleich zu einer Prophylaxedauer von weniger als 100 Tagen. Ein Vorteil in der Ausweitung der Valganciclovir-Prophylaxe auf 200 Tage ist auch im Transplantatüberleben nicht aufzuzeigen. Das Patientenüberleben wird in zwei hCMV-Risikokonstellationen [(D-/R-), (D-/R+)] signifikant unter einer 100-tägigen Prophylaxe mit Valganciclovir verlängert. Aus diesen Ergebnissen schlussfolgernd sollten gemäß der standardisierten Handlungsempfehlungen (SOP) zur hCMV-Prophylaxe nach einer Nierentransplantation am Universitätsklinikum Leipzig, Patienten, die selbst vor der Transplantation hCMV-IgG-positiv sind [(D+/R+), (D-/R+)] oder ein hCMV-positives Spenderorgan erhalten [(D+/R+), (D+/R-)] eine 100-tägige hCMV-Prophylaxe mit Valganciclovir erhalten. Ein präemptives Konzept ist für Patienten vorgesehen, die selbst vor der Transplantation hCMV-negativ sind und ein hCMV-negatives Spenderorgan erhalten. Zur Verhinderung akuter Abstoßungen und zur Optimierung des langfristigen Transplantatüberlebens erhielten Patienten mit entsprechenden Risikofaktoren eine Induktionstherapie. Die Ergebnisse dieser Arbeit zeigen, dass die Induktionstherapie keinen Risikofaktor für eine hCMV-Infektion darstellt, sie jedoch auch keinen Vorteil im Transplantat- und Patientenüberleben erzielen kann. Somit sollte eine Induktionstherapie bei Patienten mit einem hohen immunologischen Risiko zur Verhinderung akuter Abstoßungen eingesetzt werden, wodurch die Induktionstherapie auch indirekt protektiv in Bezug auf das Risiko eine hCMV-Infektion auszubilden wirken kann. Zur langfristigen Toleranz des Nierentransplantats erhielten die Patienten verschiedene Kombinationen immunsuppressiver Medikamente. Hierbei zeigt sich, dass es im gesamten Patientenkollektiv unter einem immunsuppressiven Regime mit Azathioprin zu den signifikant wenigsten hCMV-Infektionen gekommen ist. Die Patienten mit einem m-TOR-Inhibitor weisen ein signifikant längeres hCMV-freies Überleben auf, verglichen mit Patienten, die mit MMF und Tacrolimus oder Cyclosporin A immunsupprimiert wurden. In der Untersuchung des Transplantat- und Patientenüberlebens können keine signifikanten Unterschiede zwischen den verschiedenen immunsuppressiven Regimen gefunden werden. Daraus schlussfolgernd zeigen die hier vorliegenden Ergebnisse, dass ein immunsuppressives Regime mit einem m-TOR-Inhibitor insbesondere für Patienten mit einem hohen Risiko an einer hCMV-Infektion zu erkranken im Hinblick auf ein möglichst langes Transplantat- und Patientenüberleben im besonderen Maß geeignet ist. Die erhobenen Daten der am Universitätsklinikum Leipzig nierentransplantierten Patienten legen deutlich dar, dass die hCMV-Infektion eine komplexe Erkrankung ist, die einen negativen Einfluss auf das Transplantat- und Patientenüberleben nimmt. Mit einer individuellen risikofaktorenadaptierten Induktionstherapie, Immunsuppression sowie antiviralen Prophylaxe können jedoch hCMV-Infektionen reduziert werden. Zukünftig sollte zwingend eine deutschlandweite prospektive Datenbank zur Diagnostik, Prophylaxe und Therapie der humanen Cytomegalieinfektion zur stetigen Verbesserung des Nierentransplant- und Patientenüberlebens folgen.

Cytomegalievirus (CMV) ist bei immunsupprimierten Patienten nach Organ- oder Stammzelltransplantation sowie bei Patienten mit chronisch entzündlichen Darmerkrankungen und nicht immunsupprimierten kritisch kranken Patienten ein ernstzunehmender Risikofaktor für eine manifeste CMV-Infektion. Diese ist durch eine hohe Morbidität und Letalität gekennzeichnet. Daher ist eine zeitnahe und zuverlässige Diagnosestellung für die Einleitung einer adäquaten Therapie und damit für das klinische Ergebnis der Patienten von entscheidender Bedeutung. Ziel dieser retrospektiven Arbeit war es, die diagnostischen Verfahren der endoskopischen, pathologischen und virologischen Untersuchung bezüglich ihrer Treffsicherheit bei der gastrointestinalen CMV-Infektion zu untersuchen. Dafür wurde eine quantitative molekularvirologische Nachweismethode für CMV aus formalinfixiertem, in Paraffin eingebettetem Gewebe etabliert. Des Weiteren wurde das Procedere der Entnahme und der Verteilung der Biopsien an die Institute für Virologie und Pathologie evaluiert und die Ergebnisse von endoskopischer und molekularvirologischer Untersuchung in Blut- und Gewebeproben miteinander verglichen. In einem weiteren Schritt konnte die Korrelation der molekularvirologischen mit den immunhistochemischen Ergebnissen untersucht werden. Für die genannten Fragestellungen wurde CMV spezifisches Genom aus Blutproben und Gewebeproben des Gastrointestinaltraktes von 164 Patienten im Zeitraum von Oktober 2008 bis September 2010 quantitativ ausgewertet. Insgesamt wurden 860 Gewebeproben und 2550 Plasma- und Serumproben untersucht. Basierend auf den Ergebnissen der Datenerhebung zeigt die vorliegende Arbeit, dass ohne eine Anpassung der Vorgehensweise einer von vier Fällen mit gastrointestinaler CMV-Infektion nicht diagnostiziert werden würde und es wird macht einen Vorschlag zur weiteren Optimierung des diagnostischen Procedere.

Cytomegaloviruses (CMV), the prototypical β-herpesviruses, have co-evolved with their hosts and thus acquired multiple strategies for modulation of the immune response. Viral engagement of pattern recognition receptors (PRR), such as toll-like receptors (TLRs) and cytosolic nucleic acids sensors, initiates the host immune response through activation of elaborate signalling programs. The ensuing inflammatory response is further sustained and amplified through cytokines, such as IL-1β, activating signalling pathways greatly overlapping those utilized by TLRs. The central hypothesis of this thesis is that a viral counter-measure by murine CMV (MCMV) involves specific targeting of TLR- and IL-1β-induced signalling along the MyD88 to NF-κB pathway. To test this hypothesis MCMV inhibition of IL-1β signalling was initially investigated in a fibroblast cell line. It was demonstrated that in MCMV infected cells IL-1β-induced IκBα degradation is largely inhibited. Comparison of productive and non-productive infection showed this modulation requires de-novo viral gene expression beyond the immediate early region. Further investigations utilising a ORF M45 deletion mutant identified viral gene M45 as necessary for mediating the observed modulation of IL-1β- induced IκBα degradation. To further test the hypothesis, studies were extended to include TLR stimulation in the context of bone marrow-derived macrophages (BMDM) infection. It was found that TLR7/9-induced NF-κB activation is inhibited in MCMV infected BMDM. Overall, data presented in this study demonstrate a previously unrecognised MCMV inhibition of IL-1β- and TLR7/9-induced NF-κB activation, and indicate a role for viral gene M45 in mediating this effect.

Recently, cholesterol metabolism has been shown to modulate the infection of several viruses and there is growing evidence that inflammatory response to infection also modulates lipid metabolism. However little is known about the role of inflammatory processes in modulating lipid metabolism and their consequences for the viral infection. This study investigates host-lipid viral interaction pathways using mouse cytomegalovirus, a large double-stranded DNA genome, which represents one of the few models for a natural infection of its natural host. In this study, transcriptomic and lipidomic profiling of macrophages shows that there is a specific coordinated regulation of the sterol pathways upon viral infection or treatment with IFNγ or β (but not TNFα, IL1β or IL6) resulting in the decrease of free cellular cholesterol. Furthermore, we show that pharmacological and RNAi inhibition of the sterol pathway augments protection against infection in vitro and in vivo and we identified that the prenylation branch of the sterol metabolic network was involved in the protective response. Finally, we show that genetic knock out of IFNβ results in a partial reduction while genetic knock out of Ifnar1 completely abolishes the reduction of the sterol biosynthetic activity upon infection. Overall these results support a role for part of the sterol metabolic network in protective immunity and show that type 1 IFN signalling is both necessary and sufficient for reducing the sterol metabolic network upon infection; thereby linking the sterol pathway with IFN defence responses.

No description available.

Neutrophils provide a rapid first response to invading pathogens and orchestrate the immune response. They are able to employ potent antipathogenic mechanisms such as phagocytosis, reactive oxygen species (ROS) generation, protease release from granules, and formation of neutrophil extracellular traps (NETs). Despite this, certain pathogens have evolved mechanisms to benefit from neutrophil effector functions. Human cytomegalovirus (HCMV) is a clinically important pathogen that infects the majority of the human population. Monocytes are considered the main vehicle of HCMV dissemination throughout the body, but little research has been done on its interaction with neutrophils. The virus encodes a range of immunomodulatory proteins including an IL-8 homologue that acts as a powerful neutrophil chemoattractant. Viral conservation of a protein that recruits neutrophils to the site of HCMV infection suggests that the interaction between neutrophils and HCMV provides an overall advantage to the virus, but little evidence exists so far to suggest this is the case. Here I report that human peripheral blood neutrophils exposed to a clinical strain of HCMV display a profound survival phenotype, as assessed by morphology, phosphatidylserine exposure, cell permeability, and caspase-3/7 activity. This occurs in the absence of viral gene production. Neutrophils also upregulated their release of inflammatory cytokines in response to HCMV, with higher concentrations of IL-6, IL-8, and MIP-1α detected in the secretomes of infected neutrophils. These secretomes induced monocyte chemotaxis and increased monocyte permissivity to HCMV infection, as well as augmented survival in healthy, uninfected neutrophils. These experiments were confirmed with clean HCMV after the discovery of contaminating Mycoplasma spp. in the viral inocula of the initial experiments. Mycoplasma-HCMV coinfection induced an autophagic phenotype in neutrophils, as assessed by Western blotting and qPCR of autophagy-related components. Inhibition of autophagy using 3-MA reversed a profound survival effect. The unintended inclusion of Mycoplasma spp. further led to the serendipitous discovery of yet another pathogenic ability to overcome neutrophil immune functions: contaminating Mycoplasma spp. as well as Mycoplasma pneumoniae profoundly degraded NETs. These extracellular chromatin structures were stimulated using PMA or pyocyanin, and their release was dependent on the generation of ROS: severely ROS-deficient murine bone marrow neutrophils were unable to generate NETs. However, small amounts of ROS were sufficient for NETs generation, as neutrophils from acute respiratory distress syndrome patients, including many that had attenuated ROS-responses, were still capable of NETs generation. The NETs-degradative properties of mycoplasma were confirmed by fluorescence confocal and scanning electron microscopy, as well as spectrophotometry and agarose gel electrophoresis. This study demonstrates that two pervasive pathogens, HCMV and M. pneumoniae, both frequently found in coinfections in clinical contexts, are able to overcome neutrophil antipathogenic mechanisms to potentially enhance pathogen dissemination. These data provide not only a novel example of manipulation of an anti-viral response in a cell not productively infected, but also a novel example of pathogenic NETs degradation. These findings may have implications on our understanding of mycoplasma and HCMV pathogenesis and provide new targets for the generation of therapies.

The relationship between human cytomegalovirus (HCMV) infections and accelerated immune senescence is controversial. Whereas some studies reported a CMV-associated impaired capacity to control heterologous infections at old age, other studies could not confirm this. We hypothesized that these discrepancies might relate to the variability in the infectious dose of CMV occurring in real life. Here, we investigated the influence of persistent CMV infection on immune perturbations and specifically addressed the role of the infectious dose on the contribution of CMV to accelerated immune senescence. We show in experimental mouse models that the degree of mouse CMV (MCMV)-specific memory CD8(+) T cell accumulation and the phenotypic T cell profile are directly influenced by the infectious dose, and data on HCMV-specific T cells indicate a similar connection. Detailed cluster analysis of the memory CD8(+) T cell development showed that high-dose infection causes a differentiation pathway that progresses faster throughout the life span of the host, suggesting a virus-host balance that is influenced by aging and infectious dose. Importantly, short-term MCMV infection in adult mice is not disadvantageous for heterologous superinfection with lymphocytic choriomeningitis virus (LCMV). However, following long-term CMV infection the strength of the CD8(+) T cell immunity to LCMV superinfection was affected by the initial CMV infectious dose, wherein a high infectious dose was found to be a prerequisite for impaired heterologous immunity. Altogether our results underscore the importance of stratification based on the size and differentiation of the CMV-specific memory T cell pools for the impact on immune senescence, and indicate that reduction of the latent/lytic viral load can be beneficial to diminish CMV-associated immune senescence.

Orientadores: Gabriel Hessel, Sandra Cecilia Botelho Costa / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-07T22:25:30Z (GMT). No. of bitstreams: 1 Brandao_MariaAngelaBellomo_D.pdf: 1559118 bytes, checksum: 46a37bd9b6548c781556e1780d459ae7 (MD5) Previous issue date: 2006 / Resumo: A colestase neonatal pode ser a manifestação inicial de um grupo muito heterogêneo de doenças. O citomegalovírus (CMV) está entre as possíveis etiologias e os critérios para o diagnóstico não estão ainda definidos. A freqüência do CMV como causa de colestase intra-hepática (CIH) varia em função do método utilizado para o diagnóstico. O objetivo do presente estudo foi estabelecer a freqüência do CMV na colestase neonatal intra-hepática por meio dos seguintes métodos: sorologia para CMV (IgM-ELISA), N-PCR e imunohistoquímica no fragmento da biópsia hepática parafinada, e indicadores de histologia (célula de inclusão citomegálica e microabscesso) e verificar a concordância entre os métodos diagnósticos citados. Participaram do estudo 101 pacientes com o diagnóstico de CIH e que realizaram biópsia hepática . A idade dos pacientes na 1ª consulta variou de 13 dias a 7 meses, com mediana de 1 mês e 21 dias. Para determinar a freqüência da infecção por citomegalovírus foram calculados os valores de sensibilidade, especificidade, valor preditivo positivo, valor preditivo negativo e acurácia, considerando o método de N-PCR como referência. Para verificar a concordância entre os métodos, foi calculado o coeficiente kappa. A freqüência de positividade para o CMV por meio da sorologia foi de 8% (5/62), por PCR foi também de 8% (6/77), por imunohistoquímica foi de 2% (1/44). Pela avaliação histológica, nenhum em 84 pacientes apresentava células de inclusão citomegálica e 17/84 (20%) apresentavam microabscesso. A sensibilidade da sorologia em relação à N-PCR foi nula, a especificidade de 88,89%, o valor preditivo positivo nulo, o valor preditivo negativo de 90,91% e a acurácia foi de 81,63. Na pesquisa de microabscessos a sensibilidade foi de 50% em relação à N-PCR, a especificidade de 77,05%, o valor preditivo positivo de 17,65%, o valor preditivo negativo de 94% e a acurácia de 74,63%. Não houve concordância para a pesquisa de CMV entre os métodos de sorologia (ELISA-IgM) e N-PCR e fraca concordância entre os demais métodos isolados ou agrupados. Conclusões: 1. A freqüência de positividade para o citomegalovírus variou de 2% a 20% dependendo do método diagnóstico empregado, 2. Não houve concordância entre os métodos, 3. Não foram encontradas células de inclusão citomegálica e a imunohistoquímica foi positiva em apenas 1/44 casos, 4. A sorologia (ELISA-IgM) e a presença de microabscessos não foram sensíveis em relação à N-PCR para a determinação do diagnóstico de CMV na CIH, mas foram úteis, quando negativos, para predizer resultado negativo da N-PCR. Palavras chaves: lactente, hepatite / Abstract: A heterogeneous group of diseases may present initially as neonatal cholestasis, a syndrome made up of jaundice, coluria and fecal hypo or acolia. Cytomegalovirus (CMV) is one of the most common causes of neonatal intrahepatic cholestasis (IHC), but the CMV best diagnostic criteria is not yet established since the positivity of different diagnostic tests varies considerably. The aim of this study was to determine the CMV frequency in neonatal intrahepatic cholestasis and to compare results of different diagnostic tests: IgM by ELISA test, IHQ and PCR in paraffin-embedded hepatic biopsy sample and a review of liver histological features (liver microabscess and cytomegalic inclusion cells). The study has included 101 patients neonatal IHC patients who had been submitted to a liver biopsy during investigation. Median age at the first medical visit was 1 month and 21 days (13 days to 7 months). Sensibility, specificity, negative predictive value, positive predictive value and accuracy were calculated of each test in relation to N-PCR. To analyze concordance among laboratorial methods Kappa (?) coefficient was calculated. Frequency of CMV positive tests: CMV ELISA - IgM was positive in 5/62 (8%), N-PCR in 6/77 (8%), and IHQ in 1/44 (2%). Liver histological features showed 0/84 cytomegalic inclusion cells and 17/84 (20%) liver microabscess. Sensibility of serology was null, specificity was 88,98%, negative predictive value was 90,91%, positive predictive value was null, and accuracy was 81,63%. Sensibility of searching for microabscess was 50%, specificity was 77,05%, negative predictive value was 17,65%, positive predictive value was null94%, and accuracy was 74,63%.There was no concordance between ELISA-IgM and N-PCR (Value of ? =-0, 1) and weak concordance between other methods when considered as a group or individually. Conclusions: 1. Frequency of CMV varied of 2% to 20%, according the diagnostic test. 2. There was no concordance among tests. 3. Searching for inclusion cells was null and IHQ was 2%( 1/44). 4. ELISA-IgM and microabscess sensitivity were poor in relation to N-PCR, but if these methods are negative, probably N-PCR will be negative too. Key words: hepatitis, infant / Doutorado / Pediatria / Doutor em Saude da Criança e do Adolescente