Biological material is a common type of evidence found at a crime scene, and body fluid identification is an essential process in crime scene investigation. One of the most common types of body fluids found is blood. After a stain has been presumptively identified as blood through the use of a colorimetric chemical test, additional testing may be necessary to better characterize the stain. SERATEC PMB Test is a relatively new lateral flow immunochromatographic assay that targets human hemoglobin and D-dimer simultaneously in order to distinguish peripheral blood and menstrual blood at the same time. Elevated levels of D-dimer, a fibrin degradation product, are found in menstrual blood, thrombosis formation and as part of the postmortem process. A previous study investigated levels of D-dimer in menstrual, peripheral and postmortem blood using the SERATEC PMB Test. In this study, all postmortem blood samples showed positive results for both hemoglobin and D-dimer; all peripheral bloodstain samples from living individuals showed positive results for hemoglobin detection, and negative results for D-dimer detection; and most menstrual bloodstain samples showed positive D-dimer results. The results suggest that this assay could be considered a presumptive test for both postmortem blood and menstrual blood. However, as D-dimer concentrations vary between individuals, additional testing is necessary to conclusively distinguish postmortem blood, menstrual blood and peripheral blood from living individuals with especially high D-dimer levels.

A brucelose causada pela B. canis constitui uma infecção sistêmica e zoonótica que acomete principalmente os cães, causando problemas reprodutivos. O diagnóstico da infecção é difícil, sendo necessária a associação do diagnóstico clínico aos métodos laboratoriais diretos e indiretos para sua confirmação. Um dos problemas relativos ao diagnóstico laboratorial está relacionado à ausência de marcadores que possibilitem a identificação acurada de cães em ausência de bacteremia. A partir do exposto, foi realizado um estudo com o objetivo de avaliar o desempenho de testes laboratoriais diretos e indiretos como marcadores da infecção por B. canis em cães e determinar a combinação de testes que possibilite o diagnóstico da brucelose canina com maiores valores de sensibilidade e especificidade. Foram coletadas amostras de sangue, soro e aspirado de linfonodo de 92 cães, sem distinção de sexo, idade ou raça, incluindo cães reprodutores e não reprodutores. A hemocultura e a reação em cadeia pela polimerase (PCR) foram previamente realizadas nas amostras de sangue e, com base nos resultados, os 92 animais foram divididos em dois grupos: infectados (n=37) e não infectados (n= 55). Em seguida, as amostras de aspirado de linfonodo foram submetidas à PCR e ao cultivo microbiológico e as amostras de soro foram testadas por um ensaio imunocromatográfico (EIC). A partir dos resultados obtidos, a sensibilidade e especificidade diagnóstica dos testes foram calculadas utilizando os grupos infectados e não infectados, respectivamente. O coeficiente Kappa foi usado para calcular a concordância entres os testes laboratoriais e suas combinações. A proporção de resultados positivos foi de 40,2% (37/92) para os testes diretos em amostras de sangue, 29% (27/92) e 25% (23/92) para PCR e cultivo em amostras de aspirado de linfonodo, respectivamente, e 43% (40/92) para o EIC. A concordância entre os testes variou de moderada a quase perfeita. A sensibilidade e especificidade diagnóstica foram, respectivamente, 65% e 95% para PCR em amostras de aspirado de linfonodo, 62% e 100% para o cultivo microbiológico em amostras de aspirado de linfonodo e 92% e 89% para o EIC. A PCR em amostras de aspirados de linfonodos apresentou maior sensibilidade em relação ao cultivo aplicado a estas amostras, sendo uma alternativa ao diagnóstico microbiológico. A associação entre os testes de PCR em amostras de aspirados de linfonodos e de sangue e o EIC possibilitou um aumento da sensibilidade diagnóstica, por possibilitar a identificação de cães na ausência e na presença de bacteremia, com maior rapidez. / Brucellosis caused by B. canis is a systemic and zoonotic infection characterized by prolonged bacteremia that affects mainly dogs causing reproductive problems. The diagnosis of the infection is quite difficult, being necessary the association of the clinical diagnosis with direct and indirect laboratory methods to confirm the infection. The main drawback regarding the laboratory diagnosis relies on the lack of markers that allow the accurate identification of non bacteremic dogs. From the above, a study was carried out to evaluate the performance of direct and indirect laboratory tests as markers for B. canis infection in dogs and to determine the combination of tests that allows the diagnosis of the infection with higher values of sensitivity and specificity. Samples of blood, serum and lymph node aspirates were collected from 92 dogs, regardless the sex, age or breed, including pet and breeding dogs. All the dogs were tested using culturing and the polymerase chain reaction (PCR) in blood samples and, based on the results, they were divided into two groups: infected (n = 37) and non-infected (n = 55). Lymph node aspirates were tested through PCR and microbiological culturing, and serum samples using an immunochromatographic assay (EIC). The infected and uninfected groups were used to calculate, respectively, the sensitivity and specificity of the tests. The agreement between the tests was calculated using Kappa coefficient. The proportion of positive results was 40.2% (37/92) for the direct tests in blood samples, 29% (27/92) and 25% (23/92) for PCR and lymph node culturing, respectively, and 43% (40/92) for the EIC. The agreement between the tests ranged from moderate to near perfect. The sensitivity and specificity was, respectively, 65% and 95% for PCR in lymph node aspirates, 62% and 100% for lymph node culturing, and 92% and 89% for EIC. The PCR in lymph node aspirates showed a higher sensitivity when compared to the lymph node culturing, being an alternative to the microbiological diagnosis. The association between PCR in blood and lymph node aspirates and the EIC enabled an increased sensitivity in the diagnosis with the identification of non-bacteremic dogs more rapidly.

Cryptosporidium is a genus of apicomplexan unicellular epicellular parasite with worldwide distribution causing watery diarrhea in humans and animals. The life cycle is completed in one host, where Cryptosporidium parasitizing epithelial cells of gastrointestinal tract and in birds can cause disease of respiratory or urogenital tract. Course of disease depends on condition of immune system. For immunodeficient individuals could be life threatening. One of problems especially in developing countries is early and correct diagnostic, particularly no effective treatment currently exist. The aim of this thesis was to compare efficiency of immunochromatographic tests in samples stored under different conditions. The comparison of sensitivity and specificity of these tests with molecular and microscopic techniques was also performed. Additionaly, suitability of immunochromatographic tests for detection of active infection during prepatent period was evaluated. The theoretic part includes general information about Cryptosporidium. Its taxonomy, cycle of evolution or transmission and course of disease. Using of immunochromatographic test is also mentioned. No differences in sensitivity of used immunochromatographic tests was observed in this thesis. The detection rate for most of tests was 200 oocyst per sample. The presence of coproantigen is depend upon presence of oocysts in a sample. False negative results of immunochromatographic assays was caused by i) low concentration of oocysts in a sample (sensitivity) or ii) antibodies in used test don´t react with antigen of Cryptosporidium spp. (specificity). Results of this thesis show that combination of immunochromatographic tests and other techniques is convenient. During prepatent period is not possible to detect specific DNA, antigen or oocysts of Cryptosporidium. The active infection could not be distinguish from passage of oocysts using of immunochromatographic assays even if PCR is also used.

A brucelose causada pela B. canis constitui uma infecção sistêmica e zoonótica que acomete principalmente os cães, causando problemas reprodutivos. O diagnóstico da infecção é difícil, sendo necessária a associação do diagnóstico clínico aos métodos laboratoriais diretos e indiretos para sua confirmação. Um dos problemas relativos ao diagnóstico laboratorial está relacionado à ausência de marcadores que possibilitem a identificação acurada de cães em ausência de bacteremia. A partir do exposto, foi realizado um estudo com o objetivo de avaliar o desempenho de testes laboratoriais diretos e indiretos como marcadores da infecção por B. canis em cães e determinar a combinação de testes que possibilite o diagnóstico da brucelose canina com maiores valores de sensibilidade e especificidade. Foram coletadas amostras de sangue, soro e aspirado de linfonodo de 92 cães, sem distinção de sexo, idade ou raça, incluindo cães reprodutores e não reprodutores. A hemocultura e a reação em cadeia pela polimerase (PCR) foram previamente realizadas nas amostras de sangue e, com base nos resultados, os 92 animais foram divididos em dois grupos: infectados (n=37) e não infectados (n= 55). Em seguida, as amostras de aspirado de linfonodo foram submetidas à PCR e ao cultivo microbiológico e as amostras de soro foram testadas por um ensaio imunocromatográfico (EIC). A partir dos resultados obtidos, a sensibilidade e especificidade diagnóstica dos testes foram calculadas utilizando os grupos infectados e não infectados, respectivamente. O coeficiente Kappa foi usado para calcular a concordância entres os testes laboratoriais e suas combinações. A proporção de resultados positivos foi de 40,2% (37/92) para os testes diretos em amostras de sangue, 29% (27/92) e 25% (23/92) para PCR e cultivo em amostras de aspirado de linfonodo, respectivamente, e 43% (40/92) para o EIC. A concordância entre os testes variou de moderada a quase perfeita. A sensibilidade e especificidade diagnóstica foram, respectivamente, 65% e 95% para PCR em amostras de aspirado de linfonodo, 62% e 100% para o cultivo microbiológico em amostras de aspirado de linfonodo e 92% e 89% para o EIC. A PCR em amostras de aspirados de linfonodos apresentou maior sensibilidade em relação ao cultivo aplicado a estas amostras, sendo uma alternativa ao diagnóstico microbiológico. A associação entre os testes de PCR em amostras de aspirados de linfonodos e de sangue e o EIC possibilitou um aumento da sensibilidade diagnóstica, por possibilitar a identificação de cães na ausência e na presença de bacteremia, com maior rapidez. / Brucellosis caused by B. canis is a systemic and zoonotic infection characterized by prolonged bacteremia that affects mainly dogs causing reproductive problems. The diagnosis of the infection is quite difficult, being necessary the association of the clinical diagnosis with direct and indirect laboratory methods to confirm the infection. The main drawback regarding the laboratory diagnosis relies on the lack of markers that allow the accurate identification of non bacteremic dogs. From the above, a study was carried out to evaluate the performance of direct and indirect laboratory tests as markers for B. canis infection in dogs and to determine the combination of tests that allows the diagnosis of the infection with higher values of sensitivity and specificity. Samples of blood, serum and lymph node aspirates were collected from 92 dogs, regardless the sex, age or breed, including pet and breeding dogs. All the dogs were tested using culturing and the polymerase chain reaction (PCR) in blood samples and, based on the results, they were divided into two groups: infected (n = 37) and non-infected (n = 55). Lymph node aspirates were tested through PCR and microbiological culturing, and serum samples using an immunochromatographic assay (EIC). The infected and uninfected groups were used to calculate, respectively, the sensitivity and specificity of the tests. The agreement between the tests was calculated using Kappa coefficient. The proportion of positive results was 40.2% (37/92) for the direct tests in blood samples, 29% (27/92) and 25% (23/92) for PCR and lymph node culturing, respectively, and 43% (40/92) for the EIC. The agreement between the tests ranged from moderate to near perfect. The sensitivity and specificity was, respectively, 65% and 95% for PCR in lymph node aspirates, 62% and 100% for lymph node culturing, and 92% and 89% for EIC. The PCR in lymph node aspirates showed a higher sensitivity when compared to the lymph node culturing, being an alternative to the microbiological diagnosis. The association between PCR in blood and lymph node aspirates and the EIC enabled an increased sensitivity in the diagnosis with the identification of non-bacteremic dogs more rapidly.