Assessment of tubercle bacilli in various organs by staining and polymerase chain reaction technique

Background: Early diagnosis of tuberculosis (TB) by different clinical methods plays a major role in control of TB in early stages. The present study was done with the aim to assess and compare the efficiency of staining techniques and polymerase chain reaction (PCR) for detection of tubercle bacilli in various organs.Methods: The study included data 14,472 patients of both prospective (during March 2008 to 2009) and retrospective cases (past one year of the study period). For prospective cases the cytological material for the study was collected by fine needle aspiration cytology (FNAC). For retrospective cases of past one year were retrieved from the records. TB suspected cases were evaluated by Ziel-Nelson (ZN), Auramine-Rhodamine (A-R) staining techniques and by PCR.Results: Total 284 cases were diagnosed with TB in various organs. In them, epithelioid granuloma was seen 227 cases (79.92%) and Langhans giant cells were seen in 18 cases (6.34%). AFB positivity on ZN staining was observed in 161 cases (56.69%) and 9 cases (3.16%) showed positivity for tubercle bacilli on A-R staining. PCR was done in 20 prospective cases and total percentage of positivity by PCR was seen in 18 cases (90%).Conclusions: The total percentage positivity for detection of tubercule bacilli by PCR assay was found to be more (90%) with high sensitivity and specificity compared to ZN (56.6%) and AR staining techniques (3.16%).

COMPARISON OF PERMANENT STAINING METHODS FOR THE LABORATORY DIAGNOSIS OF TRICHOMONIASIS

Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in the world. The diagnosis is based on wet mount preparation and direct microscopy on fixed and stained clinical specimens. The aim of this study was to compare the performance of different fixing and staining techniques used in the detection of T. vaginalis in urine. The smears were fixed and submitted to different methods of permanent staining and then, the morphological aspects of the parasites were analyzed and compared. The Papanicolaou staining with ethanol as the fixative solution showed to be the best method of permanent staining. Our data suggest that staining techniques in association with wet mount examination of fresh specimens contribute to increase the sensitivity in the diagnosis of trichomoniasis.

Avaliação da punção aspirativa por agulha fina (PAAF) em cavidade oral e região de cabeça e pescoço em diferentes técnicas de coloração / Evaluation of fine needle aspiration biopsy (FNAB) in oral cavity and head and neck region with different techniques stains

O presente estudo teve como objetivo avaliar a Punção Aspirativa por Agulha Fina (PAAF) em diferentes técnicas de coloração, em lesões nodulares de cavidade oral e região de cabeça e pescoço, quanto a sua sensibilidade, especificidade e acurácia, nas colorações de Panótico, Papanicolau e Hematoxilina-Eosina . Foram selecionados 46 pacientes consecutivamente que procuraram a Clínica da Disciplina de Estomatologia Clínica da FOUSP, portadores de lesões nodulares em cavidade oral e região de cabeça e pescoço. Como critérios de inclusão foram selecionados pacientes de ambos os sexos, todas as etnias, acima dos 5 anos de idade, sem restrição de comorbidades e que foram realizadas PAAF com confirmação diagnóstica pela biópsia. Como critérios de exclusão da pesquisa estão os pacientes abaixo dos 5 anos de idade e pacientes que foram somente submetidos a PAAF sem confirmação diagnóstica pela biópsia. O material obtido pela PAAF foi enviado em 6 lâminas diferentes, corados pelo método de Panótico, Papanicolau e Hematoxilina-Eosina a um mesmo patologista apenas com o diagnóstico clínico. Após a emissão do laudo da PAAF, o laudo do anátomo patológico era emitido, servindo como padrão ouro. Após os cálculos, o resultado da sensibilidade, especificidade e acurácia para o método de coloração com o Panótico foram de 28,6%, 76%, 15,4, respectivamente, para o método de coloração com o Papanicolau foram de 71,4%, 76,7%, 23,3%, respectivamente e para o método de coloração com a Hematoxilina-Eosina foram de 82,1%, 23,3%, 28,6%, respectivamente...

The present study aimed to evaluate the Fine Needle Aspiration Biopsy in different staining techniques in nodular lesions of the oral cavity and head and neck region, as their sensitivity, specificity and accuracy, staining with Panoptic, Papanicolaou and Hematoxylin-Eosin. 46 patients who sought the Clinic of the Discipline of Clinical Stomatology at FOUSP were selected consecutively, with nodular lesions in the oral cavity and head and neck region. Inclusion criteria were patients of both sexes, all ethnicities, above 5 years-old, with no restriction of comorbidities and FNAB performed with confirmation by biopsy. Exclusion criteria were patients under 5 years-old and patients who only underwent FNAB without confirmation by biopsy. The material obtained by FNAB was sent on 6 different slides, stained by the method of Panoptic, Papanicolaou and Hematoxylin-Eosin, to the same pathologist only with the clinical diagnosis. After the final report of FNAB, the biopsy report was issued, serving as gold standard. After the calculations, the results of sensitivity, specificity and accuracy for Panoptic staining were 28.6%, 76% and 15.4%, respectively...

Padronização do diagnóstico de Blastocystis hominis por diferentes técnicas de coloração / Standardization of Blastocystis hominis diagnosis using different staining techniques

INTRODUÇÃO: O presente trabalho foi realizado no período de março a maio de 2008, com o objetivo de avaliar a eficácia de diferentes técnicas para diagnóstico de Blastocystis hominis em uma amostra da população atendida pelo Laboratório de Biomedicina da Universidade Feevale, Novo Hamburgo/RS. MÉTODOS: Foram estudadas 100 amostras de fezes de crianças e adultos, que foram coletadas e submetidas às técnicas de sedimentação espontânea (HPJ), sedimentação em formol-éter (Ritchie) e de coloração por Gram e May-Grünwald-Giemsa (MGG). RESULTADOS: A presença de Blastocystis hominis foi observada em 40 amostras, quando utilizadas técnicas de coloração (MGG e Gram), enquanto que as técnicas de sedimentação se mostraram menos eficientes (32 amostras positivas para a técnica de Ritchie e 20 amostras positivas para a técnica de HPJ). CONCLUSÕES: Nossos resultados demonstram que o HPJ foi menos eficiente que outros métodos, indicando a necessidade de se incluir na rotina do laboratório técnicas que permitam a identificação deste parasita.

INTRODUCTION: The present study was carried out from March to May 2008, with the aim of evaluating the effectiveness of different techniques for diagnosing Blastocystis hominis in a sample of the population attended at the Biomedicine Laboratory of Feevale University, Novo Hamburgo, Rio Grande do Sul. METHODS: On hundred feces samples from children and adults were evaluated. After collection, the samples were subjected to the techniques of spontaneous sedimentation (HPJ), sedimentation in formalin-ether (Ritchie) and staining by means of Gram and May-Grünwald-Giemsa (MGG). RESULTS: The presence of Blastocystis hominis was observed in 40 samples, when staining techniques were used (MGG and Gram), while sedimentation techniques were less efficient (32 positive samples using the Ritchie technique and 20 positive samples using the HPJ technique). CONCLUSIONS: Our results demonstrate that HPJ was less efficient than the other methods, thus indicating the need to include laboratory techniques that enable parasite identification on a routine basis.