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BACKGROUND: Bacterial identification in the clinical laboratory can be laborious and expensive. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a rapid and cost-effective diagnostic tool for the identification of organisms routinely found in the microbiology laboratory. The objective of this study was to demonstrate that identification of aerobic Gram-positive and Gram-negative organisms could be performed accurately and efficiently by MALDI-TOF MS and the Bruker Biotyper system without the use of time-consuming extraction methodologies. METHODS: Isolates previously recovered by routine culture and workup from clinical specimens were cultured to appropriate media, identified directly by MALDI-TOF MS, and compared to results from various biochemical identification methods. RESULTS: Using the direct-smear method, 99.5% and 98.0% of aerobic Gram-negative and Gram-positive bacteria, respectively, were identified to the genus level. At a score of ≥1.9, 97.6% Gram-negative organisms and 94.6% Gram-positive organisms were correctly identified to the species level by direct-smear method. Only 1.1% of isolates required further reflex to direct-plate extraction. The direct-smear method proved to be robust, as various growth temperatures, media, culture age, and different operators had no notable impact on the bacterial identification rate. CONCLUSION: The direct-smear method is an accurate and time-saving method for routine species-level bacterial identification.
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Bacterias Aerobias/aislamiento & purificación , Infecciones Bacterianas/diagnóstico , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Técnicas Bacteriológicas/métodos , Distribución de Chi-Cuadrado , Servicios de Laboratorio Clínico , Femenino , Humanos , Masculino , Estudios Retrospectivos , Temperatura , Factores de TiempoRESUMEN
OBJECTIVE: To compare direct smear technique with ethylenediaminetetraacetic acid (EDTA) preserved smear technique in terms of preparing bone marrow aspirate slides. METHODS: This prospective study was carried out between September 2009 and July 2012 at the Haematology/Oncology Department, King Khalid University Hospital, Riyadh, Saudi Arabia. With a standard gauge disposable bone marrow aspirate needle, 0.5 to 1.0 ml bone marrow was aspirated with a 10ml syringe. Half of the marrow was immediately transferred to an EDTA tube with gentle mixing, while slides were prepared directly from the rest of the sample in the syringe. The tube sample was used to prepare slides at the end of the procedure. A score of 1-4 was assigned to each slide depending on the quality and number of particles. RESULTS: A total of 245 bone marrow aspirate samples were evaluated related to 216 patients. Of the total, 238 (97%) samples were included in the study. The mean score for the direct smear group was 3.40±0.79 and for the EDTA smear group it was 3.34±0.75 (p=0.27), which was not statistically significant. An informal comparison of the morphological analysis of the samples did not reveal any differences. CONCLUSIONS: Bone marrow aspirate slides prepared at the end of the procedure from EDTA preserved samples were not inferior to slides prepared directly from the aspirated sample.
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Médula Ósea/patología , Técnicas de Preparación Histocitológica , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biopsia con Aguja , Ácido Edético , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Adulto JovenRESUMEN
Leishmaniasis can mimic many conditions, including hordeolum, basal cell carcinoma, and squamous cell carcinoma. The presence of kinetoplast in free-form or intramacrophage amastigotes, ensuring us to establish the microscopic diagnosis of leishmaniasis.
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INTRODUCTION: In most of the endemic areas, the detection of CL is based on searching for amastigotes using the direct smear method. Since expert microscopists are not usually available in every laboratory, false diagnoses are a disaster that happens. Therefore, the aim of current research is to evaluate the validity of the CL Detect™ Rapid Test (CDRT) for diagnosis CL in comparison to direct smear and polymerase chain reaction (PCR) methods. METHODS: A total of 70 patients with skin lesions suspected to be CL were recruited. Skin samples from the lesions were collected and used for direct microscopic examination and the PCR method. Furthermore, the skin sample was collected in accordance with the manufacturer's instructions for the CDRT-based rapid diagnostic test. RESULTS: Of 70 samples, 51 and 35 samples were positive by direct smear examination and the CDRT, respectively. The PCR showed positive results in 59 samples; 50 and 9 samples were identified as Leishmania major and Leishmania tropica, respectively. The sensitivity and specificity were calculated to be 68.6% (95% CI 54.11-80.89%) and 100% (95% CI 82.35-100%). When the results of CDRT were compared to the microscopic examinations, an agreement of 77.14% was seen between the CDRT and microscopic examination. In addition, the sensitivity and specificity were 59.32% (95% CI 45.75-71.93%) and 100% (95% CI 71.5-100%) when the CDRT was compared to PCR assay (as gold standard) and an agreement (65.71%) was found between CDRT and PCR assay. CONCLUSION: As the CDRT is simple, rapid, and does not require great proficiency, it is recommended for use in the detection of CL caused by L. major or L. tropica as a diagnostic method, especially in areas with limited access to expert microscopists.
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Leishmania major , Leishmania tropica , Leishmaniasis Cutánea , Humanos , ADN Protozoario/genética , Leishmaniasis Cutánea/diagnóstico , Leishmaniasis Cutánea/epidemiología , Leishmania tropica/genética , Leishmania major/genética , Reacción en Cadena de la Polimerasa/métodosRESUMEN
BACKGROUND & OBJECTIVE: Urine cytology is an important diagnostic method for urinary tract cancers (especially carcinomas), which is suitable for follow-up of residual urothelial tumors after surgery of malignant bladder tumors. Liquid-based cytology (LBC) was used for the first time in cervical cytology Compared to direct smear cytology (DSC), LBC reduced background elements (including cellular debris, inflammatory cells, and blood cells), provided better cell preservation, and had a higher satisfaction rate. In this study, we performed two different methods (DSC and LBC) to detect bladder lesions; also, we determined the sensitivity and specificity of these methods. METHODS: A total of 146 samples were taken from patients with suspected bladder cancer and processed for direct smear and LBC. In both methods, findings were reported according to the Paris System. Then, patients underwent cystoscopy and biopsy. Next, the accuracy of cytology methods was evaluated according to biopsy reports. The sensitivity and specificity of these methods were also calculated. RESULTS: Credit indices obtained for the direct smear method included sensitivity (62.5%), specificity (89%), positive predictive value (89.5%), and negative predictive value (91.5%). For LBC methods, credit indices included sensitivity (85.7%), specificity (99%), positive predictive value (96%), and negative predictive value (96%). Agreement between the two methods was statistically significant (P<0.000) in negative biopsies but not in positive biopsies (P>0.05). CONCLUSION: This study showed that LBC has higher sensitivity and specificity than the direct smear.
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BACKGROUND: Metastatic breast carcinoma (mBC) is frequently encountered and may be challenging to diagnose as the tumor cells can morphologically resemble carcinomas of other primary origins. An additional challenge is that direct smears are often the only sample type available for immunostaining studies in cytology. Trichorhinophalangeal syndrome GATA-binding type 1 1 (TRPS1) is a highly sensitive marker for BC compared to the commonly used marker GATA3, especially in triple-negative BC (TNBC), in histologic samples. However, its sensitivity and specificity in mBC and other GATA3-positive tumors have not been studied. METHODS: The authors identified the following cytology cases: 37 GATA3-positive mBC cases and 19 available cases that were deemed mBC but were GATA3-negative during the original case workup and five cases of each of eight epithelioid entities known to have high rates of GATA3 positivity and commonly seen in cytology practice. Immunostainings of TRPS1 and GATA3 were performed on the chosen smears following standard protocols. RESULTS: TRPS1 was positive in all 37 GATA3-positive mBC cases and in 18 of the 19 GATA3-negative mBC cases. TRPS1 was negative in all five of the seven frequently GATA3-positive epithelioid entities, with the exception of salivary duct carcinomas where GATA3 was positive in a rate ranging 60%-100% among them. CONCLUSIONS: TRPS1 is as sensitive as GATA3 in GATA3-positive mBC and is more sensitive than GATA3 in TNBC. TRPS1 is negative in most GATA3-positive nonbreast tumors. Thus, the combination of TRPS1 and GATA3 could be used to differentiate breast primary from others in most situations.
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Neoplasias de la Mama , Carcinoma , Humanos , Femenino , Proyectos Piloto , Biomarcadores de Tumor , Factor de Transcripción GATA3 , Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/patología , Proteínas RepresorasRESUMEN
Conventional diagnostic techniques using manual methods for stool examination have important limitations. Hence there is a need for improved technologies in routine clinical practice. This study aimed to compare detection rates, agreements, and diagnostic performances for stool examinations in all parameters of the complete filtration method using the Sciendox Feces Analysis System-50 automated feces analyzer with three manual methods, the direct smear, Kato's thick smear, and formalin ethyl concentration techniques. The 252 routine stool samples were examined for parasites, white blood cells (WBCs), red blood cells (RBCs), fat globules, and yeast cells using the four methods indicated above, and the complete filtration detection rates, Cohen's kappa (κ), and diagnostic performances were evaluated and compared. The detection rates of RBCs, fat globules, and yeast cells examined by the complete filtration automated method were comparable to the manual methods, but the detection rates of parasites and WBCs were significantly lower. Most methods detected the same seven parasite species, Ascaris lumbricoides, hookworm, Trichuris trichiura, Strongyloides stercoralis, Entamoeba histolytica/dispar, Blastocystis spp., and Giardia intestinalis. Pairwise agreements between the complete filtration and other methods were good to very good for all parameters showing κ values of 0.74 to 0.89. The diagnostic performances against the combined results showed complete filtration method sensitivities of 70%, 81.82%, 77.27%, 100%, and 95% for parasites, WBCs, RBCs, fat globules, and yeast cells, respectively, while the complete filtration negative predictive values (NPVs) and accuracies showed higher than 95% for all parameters. The complete filtration method using the automated feces analyzer showed high NPVs and accuracies, and good agreements with the three tested manual methods for stool examination in all parameters.
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Heces/parasitología , Parasitosis Intestinales/diagnóstico , Parasitosis Intestinales/parasitología , Parásitos/clasificación , Parásitos/aislamiento & purificación , Ancylostomatoidea/aislamiento & purificación , Animales , Ascaris lumbricoides/aislamiento & purificación , Blastocystis/aislamiento & purificación , Entamoeba histolytica/aislamiento & purificación , Filtración , Giardia lamblia/aislamiento & purificación , Humanos , Sensibilidad y Especificidad , Strongyloides stercoralis/aislamiento & purificación , Trichuris/aislamiento & purificaciónRESUMEN
Diagnosing EPTB is difficult as various clinical presentations are seen with paucibacillary nature of the disease. Maurya AK et al. (J Postgrad Med 58(3):185-189) AFB smear hasn't been much effective in diagnosing of EPTB. Tuberculosis of head and neck diagnosis can be missed due to large number of smear negative cases, leading false negative cases increase. CBNAAT is cartridge-based nucleic acid amplification test which detects the presence of TB bacilli this study we have examined the effectiveness of CBNAAT for diagnosing EPTB with cervical lymphadenitis and compared with Sputum AFB Microscopy smear. Aims and objective 1.To study the role of cartridge-based nucleic acid amplification test (CBNAAT) in early diagnosis of extra pulmonary tuberculosis. 2 Comparative analysis of CBNAAT with AFB smears in EPTB. Type of Study Descriptive retrospective observational study was done at the department of ENT and Head & surgery, A.B.V. Govt medical college, Vidisha, M.P, India. Result Out of 184 patients 43 samples were AFB smear positive and 141 were negative. In CBNAAT 88 out of 184 and 96 were found to be negative.32 patients were negative on Sputum microscopy AFB analysis but found to be positive on CBNAAT. Sensitivity of CBNAAT was 86% AFB 64%, Specificity of CBNAAT, AFB is 91% and 89% respectively. Our study finds that CBNAAT is much more effective in diagnosis of EPTB with cervical lymphadenitis when compared to AFB smear microscopy alone.
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INTRODUCTION: Gastric-type mucinous carcinoma (GAS) of the uterine cervix is an adenocarcinoma subtype with a gastric phenotype that poses diagnostic pitfalls in cervical screening cytology because of its blunt morphologic atypia and the limited utility of human papillomavirus testing and ancillary immunochemical staining. Despite the recent widespread uptake of liquid-based cytology (LBC) systems, the cytomorphological features of GAS in LBC samples and the differential features between GAS and usual-type endocervical adenocarcinoma (UEA) remain unclear. METHODS: Eight GAS cases, all of which were surgically treated following histological confirmation, were examined. Direct Papanicolaou-stained smears and LBC samples were reviewed and compared with 10 UEA cases as controls. Featured cytomorphological findings were as follows: background (mucinous, inflammatory, or necrotic), cell crowding (size of neoplastic cell clusters), cytoplasm (golden mucin and cell border), and nuclei (nuclear chromatin and nucleoli). RESULTS: Of 18 adenocarcinomas, 16 were detected against a non-mucinous background in LBC samples, most of which were accompanied by mild to moderate inflammation. Clusters comprising >300 neoplastic cells were identified in both GAS and UEA in conventional smears (CSs), while no LBC samples harboured clusters as large as these. Cell borders of GAS were more distinct than those of UEA in CSs (p < 0.001), although fewer populations of neoplastic clusters revealed distinct cell borders in both GAS and UEA in LBC samples. Three of 8 and 2 of 8 GAS cases had golden mucin in CSs and in LBC samples, respectively, which was not detected in UEA at all. Nucleoli against fine nuclear chromatin were more pronounced in GAS than in UEA on CS (p = 0.03), although the difference between GAS and UEA was not apparent in LBC samples. DISCUSSION/CONCLUSION: This study demonstrated that the diagnostic clues to detect GAS using the conventional approach, namely distinct cell borders and prominent nucleoli, are not useful for excluding UEA in LBC samples. Conventional cervical smears may indicate a diagnosis of GAS; however, specific high-risk HPV detection approaches, such as HPV test or immunocytochemical p16/Ki-67 dual staining, are desirable to differentiate GAS from UEA in the setting of LBC with ambiguous cytomorphological features.
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Adenocarcinoma in Situ/patología , Adenocarcinoma Mucinoso/patología , Neoplasias del Cuello Uterino/patología , Adenocarcinoma in Situ/cirugía , Adenocarcinoma Mucinoso/cirugía , Adulto , Anciano , Femenino , Humanos , Biopsia Líquida , Persona de Mediana Edad , Prueba de Papanicolaou , Valor Predictivo de las Pruebas , Reproducibilidad de los Resultados , Neoplasias del Cuello Uterino/cirugía , Frotis Vaginal , Adulto JovenRESUMEN
BACKGROUND: Intestinal schistosomiasis is still a public health problem in Burundi. Since 2008, annual mass drug administration with praziquantel has been rolled out in 11 endemic districts. The national programme relies on school-based surveys with kato-katz to monitor the impact of mass drug administration. We explored whether routine data on intestinal schistosomiasis as determined by direct fecal smears at health centre level could be used. METHODS: From the Burundian National Health Information System, we collected routine incidence data on intestinal schistosomiasis as determined by direct smear examination in all 45 sanitary districts during 2011-2015. A temporal trends analysis was performed using a mixed negative binomial regression. Sanitary districts with mass drug administration campaigns with praziquantel (n = 11) were compared with those without (n = 34). In addition, prevalence data on intestinal schistosomiasis based on kato-katz results from a school-based national mapping in 2014 were compared with the incidence data in health centres based on direct smear results, in the same 45 sanitary districts. RESULTS: In the 11 sanitary districts applying mass drug administration with praziquantel, the incidence rate decreased significantly for the years 2014 (ß2014 = - 0.826, P = 0.010) and 2015 (ß2015 = - 1.294, P < 0.001) and for the five-year period (ß = - 0.286, P < 0.001), whereas in the 34 districts where mass drug administration was not delivered, there was no significant decrease over time (ß = - 0.087, P = 0.219). In most of the 45 sanitary districts, the low prevalence based on kato-katz in school children was confirmed by low incidence rates based on direct smears in the health centres. CONCLUSIONS: National Health Information System surveillance data, based on routinely collected direct smear results at health centre level, may be able to monitor the impact of mass drug administration with praziquantel on intestinal schistosomiasis in Burundi. Control and elimination of intestinal schistosomiasis call for integration of adequate diagnosis and treatment into routine activities of primary health care facilities, as recommended by the World Health Organization since more than 20 years. When moving towards elimination, more sensitive tests, such as the point-of-care circulating cathodic antigen assay are desirable.
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Instituciones de Atención Ambulatoria/estadística & datos numéricos , Antihelmínticos/uso terapéutico , Administración Masiva de Medicamentos , Praziquantel/uso terapéutico , Esquistosomiasis/diagnóstico , Esquistosomiasis/tratamiento farmacológico , Adolescente , Burundi/epidemiología , Niño , Preescolar , Heces/parasitología , Humanos , Vigilancia de la Población/métodos , Prevalencia , Esquistosomiasis/epidemiología , Instituciones Académicas/estadística & datos numéricos , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: The role of Cartridge based Nucleic Acid Amplification test (CBNAAT) in the diagnosis of lymphnode TB which helps in reducing the mortality and morbidity by early identification and initiating treatment at the earliest. Also helps in identify the drug resistance among tubercular lymphnodes cases. PATIENTS AND METHODS: A prospective clinical study was performed in 101 suspected lymph node tuberculosis patients. The results of FNAC and/or excision biopsy of lymphnode samples obtained by CBNAAT were compared with direct smear microscopy for AFB bacilli, cytology and their combination considering AFB culture as gold standard. RESULTS: A total of 101 patients were evaluated of which 74 subjects (73.3%) were CBNAAT positive for TB. Among the CBNAAT positive cases, 57 were aged above 16 years, 38 were females, equal number (37) had single and multiple lymphnodes, 46 had less than 1cm size lymphnodes, 69 had lymphnode in neck region, 65 had chest X-ray normal. Among CBNAAT positive 74 subjects, 53 subjects (71.6%) were positive for AFB direct smear, 64 subjects (86.48%) were cytology consistent with TB and their combination were positive for TB in 71 subjects (95.94%) and 71 subjects (95.94%) were positive by AFB culture and 3 cases (0.04%) showed Rifampicin resistance. CONCLUSION: CBNAAT is a rapid diagnostic tool having sensitivity of 93.42% with specificity of 86.96% and positive predictive value of 95.95% and having comparable results with AFB culture and more sensitive than other investigation procedures. Thus it can be a rule in test for lymphnode TB.
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Mycobacterium tuberculosis/genética , Tuberculosis Ganglionar/diagnóstico , Adolescente , Adulto , Biopsia , Femenino , Humanos , Ganglios Linfáticos/microbiología , Masculino , Cuello , Técnicas de Amplificación de Ácido Nucleico , Valor Predictivo de las Pruebas , Estudios Prospectivos , Tórax , Tuberculosis Ganglionar/microbiología , Tuberculosis Ganglionar/patología , Adulto JovenRESUMEN
This review is focused on the challenges in standardising and optimising molecular testing workflow in cytopathology. Although cytological samples yield optimal quality DNA, whose minimal amounts in most cases suffice even for multigene mutational profiling, the success of molecular testing is strongly dependent on standardised preanalytical protocols for maximising DNA yield and quality. Sample cytopreparation influences, even more, the quality of RNA and consequently the potential success of reverse transcription-PCR. Here, the educational and technical involvement of the cytopathologist as a relevant component of a multidisciplinary team, in the issues related to test request, specimen collection, fixation, processing, staining, tumour fraction enrichment, DNA quality/quantity assessment and storage conditions is discussed. In addition, the specific sample requirements related to more recent technological developments are examined, underlining the modern role of the cytopathologist, whose continuous education is crucial to meet the opportunities of molecular medicine.
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Técnicas Citológicas , Análisis Mutacional de ADN/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , HumanosRESUMEN
This paper reports a case of Ancylostoma duodenale parasitized in the hepatic flexure of colon and the case was misdiagnosed at the beginning. The causes of misdiagnosis are analyzed and the laboratory examination methods of hookworm are summarized.
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Anquilostomiasis/diagnóstico , Colon Ascendente/parasitología , Ancylostoma , Animales , Errores Diagnósticos , HumanosRESUMEN
BACKGROUND AND AIMS: A nanometer silicon membrane sandwich cup system was self-designed. It could concentrate the bacilli via 0.45-µm microporous filter membrane and semi-automate the acid-fast bacilli (AFB) by a bacteria-staining machine. The aim of the study was to assess the clinical value of our self-designed system for diagnosing tuberculosis (TB). METHODS: A total of 1993 sputum specimens obtained from patients with confirmed or suspected TB were subjected to direct or concentrated specimens smear at XiangYa Hospital, Central South University between May 2012 and February 2013. In addition, all the specimens were also inoculated into Lowenstein-Jensen (L-J) media, and culture results were considered as the gold standard for calculating sensitivity and specificity. RESULTS: Compared with direct smear examination, an increased density of red stained bacilli was observed in the self-designed nanometer silicon membrane sandwich cup analysis under the microscope. The positive rate of the self-designed analysis was significantly higher than that of direct AFB smear [10.9% (217/1993) vs 6.2% (123/1993), P < 0.05]. The sensitivity of the self-designed system increased (97.3% vs 55.2%, P < 0.05) without a loss of specificity (100% vs 100%) for identifying positive TB cases compared with the direct smear method. CONCLUSION: The self-designed nanometer silicon membrane sandwich cup and semi-automatic bacteria-staining machine could more efficiently and rapidly detect the AFB in respiratory specimens than direct microscopy. This is a novel and safe examination, and it may replace direct smear examination for the diagnosis of patients with TB.
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Técnicas Bacteriológicas/instrumentación , Mycobacterium tuberculosis/aislamiento & purificación , Tuberculosis/diagnóstico , Técnicas Bacteriológicas/métodos , Femenino , Humanos , Masculino , Microscopía/métodos , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad , Silicio , Esputo/microbiologíaRESUMEN
BACKGROUND: In the era of personalized medicine, requests for molecular testing of specimens obtained with minimally invasive procedures such as fine-needle aspiration have been increasing. Although cell blocks (CBs) are the recommended specimens for molecular testing, their performance has not been well analyzed. The objective of this study was to assess the frequency and types of samples deemed unsatisfactory for molecular testing (quantity not sufficient [QNS]). METHODS: One year after the implementation of careful monitoring of QNS cases, cases submitted for lung cancer molecular testing were analyzed for the QNS rate. When the cases were rejected for the inadequacy of CBs of cytology specimens, air-dried, Diff-Quik (DQ)-stained smears were reviewed and used if they were adequate. The QNS rates were compared across 4 specimen categories: large resection, small biopsy, CB alone, and CB with DQ smears. RESULTS: One hundred seventy-six cases were studied, and 45 (25.6%) were unsatisfactory. Only 1 of 73 large resection specimens was rejected because of decalcification. The QNS rate for small biopsy specimens was 35.9% (28 of 78), whereas 64% (16 of 25) of cytology cases ordered on CBs were rejected. In combination with DQ smears, the QNS rate of cytology specimens was 32% (8 of 25), which was a significant improvement over CBs only (P = .024) and was not significantly different from the QNS rate for small biopsies (P = .671). CONCLUSIONS: The utilization of DQ-stained smears for molecular testing improves the adequacy of cytologic samples and provides a minimally invasive alternative to surgical biopsy when molecular analysis of tumor material is necessary.
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Colorantes Azulados , Pruebas Genéticas/métodos , Neoplasias Pulmonares/patología , Azul de Metileno , Manejo de Especímenes/métodos , Xantenos , Adulto , Anciano , Anciano de 80 o más Años , Biopsia con Aguja , Estudios de Cohortes , Citodiagnóstico/métodos , Análisis Mutacional de ADN , Receptores ErbB/genética , Receptores ErbB/metabolismo , Femenino , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/cirugía , Masculino , Persona de Mediana Edad , Evaluación de Necesidades , Estudios Retrospectivos , Adhesión del Tejido/métodosRESUMEN
BACKGROUND: Cutaneous Leishmaniasis (CL) is a parasitic skin disease. Diagnosis primarily is based on clinical signs and microscopic observation of parasite on direct stained smears or tissue sections. Sensitivity of direct smear is not as high as molecular methods. The aim of this study was to identify and characterize Leishmania species among the negative direct smears obtained from skin ulcers suspected to CL by PCR method. METHODS: Among 81 patients with suspicious skin lesions to CL referred to the Parasitology lab, negative Giemsa stained smears were collected. DNA extraction performed by scraping stained smears, then PCR was performed. RESULTS: Among the DNA extracted from smears, L. tropica was isolated from 9 (11.1%) of the smears and L.major was not isolated from any samples. CONCLUSION: Direct microscopy on stained smears for diagnosis of leishmaniasis is not enough accurate. PCR is recommended for clinically suspected lesions with negative result of direct smear.
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Merkel cell carcinoma represents a highly aggressive cutaneous malignancy characterized by regional recurrences, lymph node metastases, distant metastases, and high mortality. As the cytomorphology of Merkel cell carcinoma can be mimicked by other malignancies, especially lymphoma and pulmonary small cell carcinoma, immunocytochemistry is often useful in confirming the diagnosis. Cell blocks, which are traditionally utilized for immunocytochemistry, occasionally exhibit insufficient cellularity. Hence, we prospectively investigated the application of CK20 immunocytochemistry to air-dried, unstained direct smears in the diagnosis of Merkel cell carcinoma fine needle aspirates (FNAs). Eight consecutive FNAs of Merkel cell carcinoma were prospectively examined in this series; seven (88%) cases exhibited immunoreactivity for CK20 in the tumor cells. The one CK20-negative Merkel cell carcinoma was immunoreactive for synaptophysin and CD56. This immunophenotype was identical to that of the original primary tumor. For comparison, air-dried direct smears prepared from three pulmonary small cell carcinoma FNAs were examined by CK20 immunocytochemistry. In all cases, no CK20 immunoreactivity was seen in any of the tumor cells. In conclusion, direct smears represent a feasible and robust source of cellular material for immunocytochemical studies to diagnose Merkel cell carcinoma. This methodology allows the cytologist to confirm on site that material for diagnostic immunocytochemistry is present thereby serving as a safeguard in instances where insufficient cell block cellularity is anticipated or encountered.
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Carcinoma de Células de Merkel/secundario , Neoplasias Cutáneas/patología , Biopsia con Aguja Fina , Carcinoma de Células de Merkel/diagnóstico , Carcinoma de Células de Merkel/metabolismo , Humanos , Inmunohistoquímica , Queratina-20/metabolismo , Metástasis Linfática , Estudios Prospectivos , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/metabolismo , Sinaptofisina/metabolismoRESUMEN
BACKGROUND: Rearrangements involving the anaplastic lymphoma kinase (ALK) gene are present in approximately 5% of lung adenocarcinomas. Crizotinib is approved for the treatment of lung adenocarcinomas harboring ALK rearrangements. Patients with advanced stage lung cancer are not candidates for surgical resection of their primary tumors. For these patients, cytologic specimens often represent the only diagnostic tissue available. Cell blocks (CBs) are routinely used for molecular studies; however, insufficient CB cellularity can impede the performance of these assays. METHODS: Thirty-two cytology cases of lung adenocarcinomas were analyzed by fluorescence in situ hybridization (FISH) for ALK rearrangements. Diff-Quik-stained smears were examined to identify tumor cell-enriched areas that were marked using a diamond-tipped scribe. Paired ALK rearrangement FISH was performed using smears and CBs in each case. RESULTS: An ALK rearrangement was detected on direct smears and CB sections in 5 (16%) and 4 (13%), respectively, of the 32 cases studied. Concordant FISH results for smears and CBs were observed in 31 (97%) of 32 cases. In the 1 discordant case, an ALK rearrangement was detected on the direct smear but not in the CB. Reverse transcriptase-polymerase chain reaction analysis of this CB revealed the presence of an EML4-ALK rearrangement, thereby confirming a false-negative FISH result in the CB. CONCLUSIONS: Stained cytologic direct smears can be effectively used for ALK rearrangement analysis by FISH. This approach represents a useful safeguard when insufficient CB cellularity is encountered and could prevent delays in treatment in this era of precision medicine.
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Adenocarcinoma/genética , Biomarcadores de Tumor/genética , Citodiagnóstico , Reordenamiento Génico , Neoplasias Pulmonares/genética , Proteínas de Fusión Oncogénica/genética , Proteínas Tirosina Quinasas Receptoras/genética , Adenocarcinoma/diagnóstico , Quinasa de Linfoma Anaplásico , Humanos , Hibridación Fluorescente in Situ , Neoplasias Pulmonares/diagnóstico , Pronóstico , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
This paper reports a case of Ancylostoma duodenale parasitized in the hepatic flexure of colon and the case was misdiagnosed at the beginning. The causes of misdiagnosis are analyzed and the laboratory examination methods of hookworm are summarized.