Source: Masters Abstracts International, Volume: 47-06, page: 3545. Adviser: Leona K. Bartholomew. Includes bibliographical references.

A research report submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in partial fulfilment of the requirements for the degree of Master of Medicine in the branch of Haematology. Johannesburg, March 2015 / The incidence of leukaemia in South Africa is 2.5 per 100 000 and has increased due to HIV. Accurate and timeous diagnosis of leukaemia directly impacts success of patient treatment and consequent survival. Usually the Full Blood count (FBC), white blood cell (WBC) differential count and review of the peripheral blood smear alerts the clinician to the possibility of leukaemia. However the number of qualified and skilled technologists in peripheral and central laboratories is on a continual decline making the performance of the critical function of peripheral blood review a challenge. The Advia 2120 haematology analyser performs a WBC and differential count using principles of flow cytometry and the cytograms generated can be used to classify haematological malignancies through the Peroxidase and nuclear density analysis (PANDA) classification system. The presence of myeloperoxidase (MPO) activity in 3% or more leukaemic blasts confirms acute myeloid leukaemia, and enzyme activity can be detected by immunophenotypic analysis or conventional cytochemistry . Research on the comparison of the Advia 2120 and manual morphologic assessment in the classification of leukaemias is limited in the South African setting, where leukaemia often coincides with infection. The aim of this study was to determine if the FBC, differential count and cytogram assessment by the Advia 2120 using the PANDA classification is as reliable as morphologic assessment in the initial classification of haematological malignancies from peripheral blood samples when using flow cytometry as the gold standard.. 150 cases of confirmed leukaemia were collected. The diagnosis obtained from either PANDA analysis and/or morphological assessment was compared to the diagnosis obtained by immunophenotypic analysis. Secondly, the MPO activity obtained by the Advia peroxidase cytogram was compared to the MPO obtained by conventional methods of immunophenotypic analysis and/or cytochemistry. Using the PANDA analysis system, only 48% (72/150) of cases overall were accurately classified. The inaccuracy was 9.3% (14/150) and 42.7% of cases could not be classified. The positive predictive value (PPV) was 88%. The most significant finding was all of the acute Page | iv promyelocytic leukaemia (APL) cases (8/8) had a distinct pattern and were accurately classified on cytogram analysis alone. Accurate sub-classification of other types of acute myeloid leukaemia using PANDA analysis alone was inconsistent. However, the accuracy in classifying leukaemia was improved when the Advia cytogram was used in conjunction with morphological analysis, as 90% (135/150) of cases were accurately classified. The sensitivity and specificity of the peroxidase cytogram in evaluating myeloperoxidase (MPO) activity was 85% and 88.6% respectively. The agreement between cytogram peroxidase activity and the reference methods was 89.1% and the Cohen’s kappa was 76.9%. To the best of our knowledge, there is no data comparing peroxidase activity on the cytogram to other methods. In conclusion, it was shown that the routine use of the Advia cytograms in conjunction with the morphology findings provides invaluable information in the initial screening of leukaemia. In cases with indistinct morphology, the cytograms have the potential to aid in a provisional classification. The peroxidase activity from the cytogram could be used as a surrogate marker for myeloperoxidase activity in leukaemia. Moreover, a tentative diagnosis of an APL is possible by simple analysis of the cytogram resulting in earlier diagnosis which could be life-saving.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Univ., 2001. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

Source: Masters Abstracts International, Volume: 48-02, page: . Adviser: David Hewett-Emmett. Includes bibliographical references.

Orientadores: Maria Luiza Moretti, Angélica Zaninelli Schreiber / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-27T08:47:24Z (GMT). No. of bitstreams: 1 Moraes_JoseRenatode_M.pdf: 2487255 bytes, checksum: 4c5a21c13a0b57cf864dbf79cca990fd (MD5) Previous issue date: 2014 / Resumo: Introdução: os fungos do gênero Fusarium são ubíquos no meio ambiente e causam doenças em plantas e aos seres humanos. Objetivos: 1)estabelecer o protocolo para a coleta e o isolamento de Fusarium spp. no ar; 2)isolar fungos do gênero Fusarium do ar da enfermaria de Hematologia e da Unidade de Transplante de Medula Óssea (TMO) no Hospital de Clínicas (HC) da Universidade Estadual de Campinas (UNICAMP) e comparar a relação genética de isolados ambientais de Fusarium spp. com os isolados a partir de hemoculturas de rotina diagnóstica de pacientes hospitalizados durante os anos de 2002 a 2013, 3)identificar os fungos do gênero Fusarium ao nível de complexo de espécies por PCR em tempo real, e ao nível de espécie pelo sequenciamento de fragmento do gene EF1?. Metodologia: foram colhidas amostras de ar nas unidades de TMO (que possui controle de ar através de filtros HEPA e fluxo de ar de pressão positiva) e na Hematologia (unidade sem controle do ar) do HC. De 2006 a 2013 foram identificados 18 isolados de Fusarium spp. em culturas de sangue. A coleta de ar foi realizada através do amostrador de ar BioSamp modelo MBS 1000D (YotsubishiCorp Japão). Um meio de cultura seletivo para Fusarium (Agar Komada com modificações) foi usado na placa de Petri dentro do amostrador de ar. O volume de ar de 1000 mL e 500 mL foram aspirados da Hematologia e enfermarias de TMO, respectivamente. Após o tempo de incubação, as colônias que cresceram nas placas foram avaliadas macroscopicamente e microscopicamente para identificação morfológica. Fusarium spp. foram identificados através de um novo ensaio de PCR em tempo real composto por dois ensaios de PCR em tempo real: um específico para o gênero Fusarium e um ensaio específico para detecção específica do complexo de espécies de Fusarium solani (FSSC). Para confirmar o complexo de espécies de Fusarium, uma região parcial do gene EF-1? foi sequenciado. O alinhamento das sequências foi realizado no programa Clustal W e a análise filogenética utilizou o programa Mega5 com o algorítimo Neighbor-Joininge Bootstrap de 1000 replicatas. Resultado: foram isoladas 108 cepas de Fusarium spp. destes 28% na unidade de TMO e 78% na Hematologia. Os complexos de espécies encontrados foram: 10% FCSC, 21% FIESC, 1% FOSC, 12% FSSC, 51% GFSC e 5% não definido. Na TMO a maior prevalência foi da espécie Fusarium solani (9/30 isolados) e na Hematologia Fusarium verticillioides (25/31 isolados). Não foi possível constatar relevância entre a temperatura e umidade do ar em relação ao aumento ou diminuição de isolados nas coletas. Nove isolados FSSC do ar e 8 de sangue apresentaram BT 99%. Conclusão. Os dados de análise filogenética sugeriram que os isolados de sangue de pacientes e ambientais foram similares sugerindo que o ambiente pôde representar uma fonte potencial de infecção para os pacientes imunodeprimidos / Abstract: Introduction:the fungus Fusarium are ubiquitous in the environment and cause diseases in plants and humans. Objectives: isolate fungi Fusarium in the air of the ward Hematology and Unit of Bone Marrow Transplantation (BMT) at the Hospital de Clinicas (HC), State University of Campinas (UNICAMP), compare the phylogenetic relationship of environmental isolates of Fusarium spp .with isolates from blood cultures of routine diagnosis of hospitalized patients during the years 2002-2013, to establish protocol for the collection and isolation of Fusarium spp. in the air, identify the fungi Fusarium species complex level by real time PCR, and to species level by sequencing the fragment EF1 ? gene. Methodology: air sample was collected in BMT units (which has control of air through HEPA filters and air flow positive pressure) and Hematology (there is not self control air inlet) HC. From 2006 to 2013, 18 samples obtained from blood cultures with the presence of Fusarium spp. were added to study the relationship between degree of molecular clinical isolates isolated from the air. The air sampling was performed using the Bio air sampler Samp MBS model 1000D (Yotsubishi Corp. Japan). A selective medium for Fusarium (Komada Agar changes by Y. Mikami, Chiba University) was used in the petri dish in the air sampler. The air volume of 1000 mL and 500 mL were aspirated Hematology and BMT wards, respectively. After the incubation time, colonies that grew on the plates were evaluated macroscopically and microscopically for morphological identification. Fusarium spp. were identified by PCR of a new system in real time according to Muraosa et al. comprising two PCR assays in real time: the specific assay Fusarium and a specific assay for specific detection of Fusarium solani (FSSC) complex. To confirm the complex of Fusarium species, a partial region of the EF-1? gene was sequenced. The alignment of sequences was performed in Clustal W program and phylogenetic analysis used Mega5 program with the neighbor-joining algorithm and bootstrap 1000 replicates. Results: 108 ceepas Fusarium spp were isolated. 28% of these isolates form the BMT unit and 78% in Hematology. Overall the species complexes ficarm divided as follows: 10% FCSC, 21% FIESC 1% FOSC, FSSC 12%, 51% and 5% GFSC nd not defined). In the BMT was higher prevalence of the species Fusariu solani (9/30 isolates) and the Hematology espéciecom largest presence is Fusarium verticillioides (25/31 isolates). Unable to find relevance between temperature and air humidity in relation to the increase or decrease in collections of isolates. Nine isolates of Fusarium solani air obtained from the same source ancestras 8 Fusarium solani isolates from blood (BT 99%). Conclusion:Fusarium spp were isolated in 108 environmental samples en all evaluated environments. Phylogenetically findings suggest equivalence between environmental isolates and isolates from the blood of patients. The collection protocol set showed satisfactory for the specific isolation of Fusarium result. The findings of Fusarium were grouped into complexes FSSC and FSSC not the technique of real-time PCR and sequencing of the gene EF1? allowed to classify the isolates in the species level / Mestrado / Clinica Medica / Mestre em Ciências