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1.
Article in English | WPRIM | ID: wpr-922566

ABSTRACT

COVID-19 virus is a causative agent of viral pandemic in human beings which specifically targets respiratory system of humans and causes viral pneumonia. This unusual viral pneumonia is rapidly spreading to all parts of the world, currently affecting about 105 million people with 2.3 million deaths. Current review described history, genomic characteristics, replication, and pathogenesis of COVID-19 with special emphasis on Nigella sativum (N. sativum) as a treatment option. N. sativum seeds are historically and religiously used over the centuries, both for prevention and treatment of different diseases. This review summarizes the potential role of N. sativum seeds against COVID-19 infection at levels of in silico, cell lines and animal models.


Subject(s)
Animals , COVID-19 , Humans , Nigella , Pandemics , Pathology, Molecular , SARS-CoV-2
2.
Oncol. (Guayaquil) ; 31(2): 93-103, 31 de agosto 2021.
Article in Spanish | LILACS | ID: biblio-1284421

ABSTRACT

Introducción: El carcinoma basocelular (CBC)es una de las neoplasias más comunes de la piel. En nuestro país, por su localización, representa una entidad patológica de gran importancia, por la radiación ultravioleta elevada, que es inversamente proporcional a la latitud geográfica en la que nos encontramos en Ecuador. El objetivo del presente trabajo es revisar las características claves que distinguen al Carcinoma basocelular, y actualizar los conocimientos, incluyendo la evidencia disponible en hallazgos histopatológicos, marcadores de inmunohistoquímica y metástasis en esta patología


Introduction: Basal cell carcinoma BCC is one of the most common skin neoplasms. In our country, because of its location, it represents a pathological entity of great importance, due to the high ultraviolet radiation, that is inversely proportional to the geographical latitude we are in Ecuador. This paper objective is to review the key features that distinguish basal cellcarcinoma, and update knowledge, including the available evidence on histopathological findings, immunohistochemical markers and metastasis in this pathology.


Introdução: Carcinoma basocelular O CBC é uma das neoplasias cutâneas mais comuns. Em nosso país, por sua localização, representa uma entidade patológica de grande importância, devido à alta radiação ultravioleta, que é inversamente proporcional à latitude geográfica em que nos encontramos no Equador. O objetivo deste artigo é revisar as principais características que distinguem o carcinoma basocelular e atualizar o conhecimento, incluindo as evidências disponíveis sobre achados histopatológicos, marcadores imunohistoquímicos e metástases nessa patologia.


Subject(s)
Humans , Adult , Immunohistochemistry , Carcinoma, Basal Cell , Skin Neoplasms , Pathology, Molecular , Neoplasm Metastasis
3.
Chinese Journal of Biotechnology ; (12): 673-679, 2021.
Article in Chinese | WPRIM | ID: wpr-878592

ABSTRACT

Nucleic acid detection technique has good sensitivity and specificity and is widely used in in vitro diagnosis, animal and plant commodity quarantine, forensic identification, and other fields. However, it is susceptible to carryover contamination during the operation and leads to false-positive results, which seriously affects the detection accuracy. Therefore, finding an effective solution to prevent and eliminate nucleic acid carryover contamination has become particularly urgent. This study compared several different methods for removing nucleic acid contamination and confirmed that sodium hypochlorite solution and PCRguard reagent could effectively eliminate nucleic acid carryover in the liquid and on surfaces of different materials. Besides, the combination of sodium hypochlorite solution and PCRguard can solve the nucleic acid aerosol contamination. This study proposes solutions for the routine prevention of carryover contamination and removal of aerosol that has occurred in molecular diagnostic laboratories.


Subject(s)
Laboratories , Nucleic Acids , Pathology, Molecular
4.
Repert.Med.Cir ; 30(3): 214-218, 2021.
Article in English, Spanish | LILACS, COLNAL | ID: biblio-1362924

ABSTRACT

La patología y la salud pública son disciplinas que se complementan en múltiples formas, desde la información que aportan mutuamente a niveles individual y poblacional, hasta la elaboración de políticas públicas en salud y la gestión de la información en los biobancos, así como la articulación para respuesta en emergencias y brotes. En revisión no sistemática resaltamos que los dos campos de mayor colaboración con la salud pública son la patología forense (comprendiendo muertes violentas y no violentas) y la patología molecular, realizando aportes significativos a la planeación de los servicios de salud, la calidad de la información epidemiológica, la salud pública basada en la evidencia que permite una mejor toma de decisiones, y la gestión de la salud comunitaria y poblacional. A partir de la revisión realizada se identificaron como puntos de mejora el uso de los sistemas de información, la necesidad de un enfoque interdisciplinario más tangible, y la urgente transformación educativa que subyace a esta colaboración.


Pathology and public health are disciplines that complement each other in many ways, from the information they provide to each other at the individual and population levels, to the development of public health policies and the management of information in biobanks, as well as articulation in responding to emergencies and outbreaks. Our non-systematic review highlights that the two most relevant fields which collaborate with public health are forensic pathology (including violent and non-violent deaths) and molecular pathology, making significant contributions to health care planning, the quality of epidemiological information, evidence-based public health that enables better decision-making, and community and population health management. This review identified the use of information systems, the need for a more tangible interdisciplinary approach, and the urgent educational transformation that underlies this collaboration, as areas for improvement.


Subject(s)
Public Health , Health Policy , Pathology, Clinical , Autopsy , Forensic Pathology , Pathology, Molecular
5.
Rev. medica electron ; 42(5): 2355-2365, sept.-oct. 2020. graf
Article in Spanish | LILACS, CUMED | ID: biblio-1144739

ABSTRACT

RESUMEN Los ependimomas surgen de las células ependimarias que revisten los ventrículos y los pasajes en el encéfalo y el centro de la médula espinal. Las células ependimarias producen líquido cefalorraquídeo. Se decidió la realización de una revisión acerca del ependimoma intracraneal teniendo en cuenta que no existe artículo nacional que trate este tema, siendo la mayoría de los trabajos consultados referentes a la misma variante histológica pero en localización espinal, cuyo objetivo es describir la características clínicas, moleculares y anatomopatológicas del ependimoma intracraneal. Se realizó la búsqueda de artículos en revistas de las bases de datos: PubMed, Scielo y EBSCO. La búsqueda se limitó a artículos con el texto completo, publicados fundamentalmente en los últimos cinco años. El ependimoma intracraneal es un tumor frecuente en la edad pediátrica, sus manifestaciones clínicas dependen de su localización, presenta una gran diversidad molecular y anatomoptológica (AU).


SUMMARY Ependymomas arise from ependymal cells that line the ventricles and passages in the brain and center of the spinal cord. Ependymal cells produce cerebrospinal fluid. It was decided to conduct a review about intracranial ependymoma taking into account that there is no national article dealing with this issue, with most of the works consulted referring to the same histological variant but in spinal location, whose objective is to describe the clinical characteristics, Molecular and pathological pathways of intracranial ependymoma. We searched articles in journals of the databases: PubMed, Scielo and EBSCO. The search was limited to articles with the full text, published mainly in the last five years. Intracranial ependymoma is a frequent tumor in the pediatric age, its clinical manifestations depend on its location, it has a great molecular and anatomoptological diversity (AU).


Subject(s)
Humans , Male , Female , Child , Ependymoma/epidemiology , Neoplasms/diagnosis , Pathology, Clinical/methods , Signs and Symptoms , Child , Ependymoma/complications , Ependymoma/diagnosis , Pathology, Molecular/methods
6.
Rev. ADM ; 77(3): 162-167, mayo-jun. 2020. ilus
Article in Spanish | LILACS | ID: biblio-1128895

ABSTRACT

Introducción: El síndrome de Gorlin-Goltz o síndrome de carcinoma de nevo basocelular es un desorden hereditario autosómico dominante que predispone principalmente a la proliferación de múltiples carcinomas basocelulares, queratoquistes odontogénicos y defectos del desarrollo, causados por la mutación del gen Patched localizado en el cromosoma 9. Presentación del caso: Se reporta un paciente con características de este síndrome, en la clínica de COMF de la UNAM. El diagnóstico fue basado en los estudios clínicos, imagenológicos y moleculares. Conclusiones: El conocimiento de esta enfermedad puede orientarnos a la sospecha diagnóstica de lesión quística o premaligna en forma oportuna, lo que permite prevenir complicaciones y brindar un tratamiento integral para así mejorar la calidad de vida de este tipo de pacientes (AU)


Introduction: Gorlin-Goltz syndrome or cell-based nevus carcinoma syndrome is an autosomal dominant inherited disorder that predisposes mainly to the proliferation of multiple basal cell carcinomas, maxillary keratocysts and developmental defects, caused by the mutation of the Patched gene located on chromosome 9. Case presentation: A patient with specific characteristics compatible with this syndrome was reported in the COMF Department of the UNAM. The diagnosis was based on clinical studies, radiology and genetic studies. Conclusions: Knowledge of this problem can guide us to the diagnostic suspicion in a timely manner, thus preventing complications, and to provide an improved integral treatment of the quality of life of this type of patients (AU)


Subject(s)
Humans , Male , Child , Carcinoma, Basal Cell , Basal Cell Nevus Syndrome , Odontogenic Cysts/surgery , Oral Manifestations , Biopsy , Histological Techniques , Pathology, Molecular , Patched-1 Receptor , Mexico
9.
Article in English | WPRIM | ID: wpr-816637

ABSTRACT

Sexually transmitted infections (STIs) are caused by the spread of pathogens via sexual activity and can cause serious complications if left untreated, regardless of their symptoms. Therefore, early diagnosis of STI is important, and molecular diagnostic methods for rapid detection and monitoring are needed. In this study, we evaluated a multiplex polymerase chain reaction (PCR) kit for simultaneously detecting 13 different bacterial, fungal, and viral microorganisms that cause STIs. The kit performance was evaluated for its sensitivity, lot-to-lot variation, and interference in detecting different pathogens. Additionally, its clinical usefulness was evaluated by estimating its sensitivity and specificity for clinical samples. The limit of detection (LOD) was 0.021–50.104 copies for each pathogen. In the tests of lot-to-lot, 100% of positive samples were detected at low concentrations and negative samples all showed negative results. This result confirms that there is no the variation of lot-to-lot. In the test for interference between pathogens, the efficiency of amplification for each pathogen was not significantly reduced and no nonspecific amplification product was formed. We tested 322 vaginal swab samples using the multiplex PCR kit and confirmed that its clinical sensitivity and specificity were 100% for all pathogens. This multiplex PCR kit can be used widely for rapid diagnosis and monitoring of STIs.


Subject(s)
Diagnosis , Early Diagnosis , Limit of Detection , Multiplex Polymerase Chain Reaction , Pathology, Molecular , Sensitivity and Specificity , Sexual Behavior , Sexually Transmitted Diseases
10.
11.
Rev. Soc. Bras. Med. Trop ; 53: e20200314, 2020. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS, SES-SP | ID: biblio-1136805

ABSTRACT

Abstract INTRODUCTION: Rapid and accurate tuberculosis detection is critical for improving patient diagnosis and decreasing tuberculosis transmission. Molecular assays can significantly increase laboratory costs; therefore, the average time and economic impact should be evaluated before implementing a new technology. The aim of this study was to evaluate the cost and average turnaround time of smear microscopy and Xpert assay at a university hospital. METHODS: The turnaround time and cost of the laboratory diagnosis of tuberculosis were calculated based on the mean cost and activity based costing (ABC). RESULTS: The average turnaround time for smear microscopy was 16.6 hours while that for Xpert was 24.1 hours. The Xpert had a mean cost of USD 17.37 with an ABC of USD 10.86, while smear microscopy had a mean cost of USD 13.31 with an ABC of USD 6.01. The sensitivity of smear microscopy was 42.9% and its specificity was 99.1%, while the Xpert assay had a sensitivity of 100% and a specificity of 96.7%. CONCLUSIONS: The Xpert assay has high accuracy; however, the turnaround time and cost of smear microscopy were lower than those of Xpert.


Subject(s)
Humans , Tuberculosis, Pulmonary/diagnosis , Biological Assay/economics , Pathology, Molecular/economics , Tuberculosis , Tuberculosis, Pulmonary/economics , Biological Assay/methods , Sensitivity and Specificity , Costs and Cost Analysis , Pathology, Molecular/methods , Microscopy , Mycobacterium tuberculosis
12.
Arch. argent. pediatr ; 117(3): 274-278, jun. 2019. ilus
Article in English, Spanish | LILACS, BINACIS | ID: biblio-1001201

ABSTRACT

El síndrome de Ehlers-Danlos es un conjunto de trastornos hereditarios del tejido conectivo, clínica y genéticamente heterogéneos, caracterizados por hiperextensibilidad cutánea, pobre cicatrización, hipermovilidad articular y friabilidad tisular. Desde 1997, se han reportado variantes poco frecuentes del síndrome, entre las cuales se incluye el de tipo cifoescoliótico, causado por mutaciones en el gen PLOD1, caracterizado por hipotonía muscular grave al nacer, cifoescoliosis grave progresiva, osteopenia, ojos frágiles y fragilidad vascular. También ha sido descrita una rara variante recesiva que compromete el gen FKBP14, con hallazgos clínicos adicionales, que incluyen retardo del desarrollo psicomotor, miopatía, hipoacusia y una proporción normal de lisil-piridinolina a hidroxilisil-piridinolina en la orina. Se presenta el primer caso de una paciente colombiana con una mutación FKBP14 c.362dupC, caracterizada por hipotonía generalizada, retardo en el desarrollo de los hitos motores gruesos, hipoacusia, cifoescoliosis progresiva temprana, hipermovilidad articular y deformidades en los pies.


Ehlers-Danlos syndrome (EDS) is a group of clinically and genetically heterogeneous inherited connective tissue disorders, characterized by skin hyperextensibility, poor wound healing, joint hypermobility and tissue friability. Since 1997 a new spectrum of novel rare EDS-variants has been described, among which is included the EDS kyphoscoliotic type, characterized by severe muscular hypotonia at birth, severe progressive kyphoscoliosis, osteopenia, fragile eyeballs and vascular fragility. This EDS variant is caused by mutations in the PLOD1 gene; however, a rare recessive variant that compromises the FKBP14 gene has been reported, with additional clinical findings that includes gross motor developmental delay, myopathy, hearing impairment and a normal ratio of lysyl pyridinoline to hydroxylysyl pyridinoline in urine. We report the first Colombian patient with a FKBP14 c.362dupC mutation, with clinical features that include generalized hypotonia, delayed gross motor milestones, hearing loss, early-onset progressive kyphoscoliosis, joint hypermobility and foot deformities.


Subject(s)
Humans , Female , Adolescent , Spinal Curvatures , Ehlers-Danlos Syndrome , Pathology, Molecular
13.
Article in English | WPRIM | ID: wpr-719474

ABSTRACT

The detection and quantification of hepatitis B virus (HBV) DNA plays an important role in diagnosing and monitoring HBV infection as well as in assessing the therapeutic response. We compared the analytical performance of a random access, fully automated HBV assay—DxN VERIS Molecular Diagnostics System (Beckman Coulter, Brea, CA, USA)—with that of Abbott RealTime HBV assay (Abbott Laboratories, Des Plaines, IL, USA). The between-day precision of the VERIS assay ranged from 0.92% (mean 4.68 log IU/mL) to 4.15% (mean 2.09 log IU/mL) for pooled sera from HBV patients. HBV DNA levels measured by the VERIS HBV assay correlated with the calculated HBV DNA levels (r²=0.9994; P < 0.0001). The lower limit of quantification was estimated as 8.76 IU/mL (Probit analysis, 95% confidence interval: 7.32–12.00 IU/mL). Passing-Bablok regression analysis showed good concordance between the VERIS and RealTime assays for 187 chronic HBV samples (y=−0.2397+0.9712x; r=0.981), as well as for 20 drug-resistant HBV genotype C positive samples (y=−0.5415+0.9954x; r=0.961). The VERIS assay demonstrated performance similar to the RealTime assay and is suitable for high-throughput HBV DNA monitoring in large hospital laboratories.


Subject(s)
DNA , Genotype , Hepatitis B virus , Hepatitis B , Hepatitis , Humans , Laboratories, Hospital , Pathology, Molecular
14.
Article in English | WPRIM | ID: wpr-741212

ABSTRACT

BACKGROUND: Recent findings in molecular pathology suggest that genetic translocation and/or overexpression of oncoproteins is important in salivary gland tumorigenesis and diagnosis. We investigated PLAG1, SOX10, and Myb protein expression in various salivary gland neoplasm tissues. METHODS: A total of 113 cases of surgically resected salivary gland neoplasms at the National Cancer Center from January 2007 to March 2017 were identified. Immunohistochemical staining of PLAG1, SOX10, and Myb in tissue samples was performed using tissue microarrays. RESULTS: Among the 113 cases, 82 (72.6%) were benign and 31 (27.4%) were malignant. PLAG1 showed nuclear staining and normal parotid gland was not stained. Among 48 cases of pleomorphic adenoma, 29 (60.4%) were positive for PLAG1. All other benign and malignant salivary gland neoplasms were PLAG1-negative. SOX10 showed nuclear staining. In normal salivary gland tissues SOX10 was expressed in cells of acinus and intercalated ducts. In benign tumors, SOX10 expression was observed in all pleomorphic adenoma (48/48), and basal cell adenoma (3/3), but not in other benign tumors. SOX10 positivity was observed in nine of 31 (29.0%) malignant tumors. Myb showed nuclear staining but was not detected in normal parotid glands. Four of 31 (12.9%) malignant tumors showed Myb positivity: three adenoid cystic carcinomas (AdCC) and one myoepithelial carcinoma with focal AdCC-like histology. CONCLUSIONS: PLAG1 expression is specific to pleomorphic adenoma. SOX10 expression is helpful to rule out excretory duct origin tumor, but its diagnostic value is relatively low. Myb is useful for diagnosing AdCC when histology is unclear in the surgical specimen.


Subject(s)
Adenoma , Adenoma, Pleomorphic , Antibody-Dependent Cell Cytotoxicity , Carcinogenesis , Carcinoma, Adenoid Cystic , Diagnosis , Immunohistochemistry , Oncogene Proteins , Oncogene Proteins v-myb , Parotid Gland , Pathology, Molecular , Salivary Gland Neoplasms , Salivary Glands , SOX Transcription Factors , Translocation, Genetic
15.
Article in Korean | WPRIM | ID: wpr-786351

ABSTRACT

Pancreas cystic neoplasm is a relatively common disease. However, its' pathologic diagnosis is not easy. The most frequent problem is low cellularity when compared to another organ cytology or biopsy material. Considering the procedure and anatomic difficulty, it is not uncommon to observe a low cellular smear or scanty volume of cells in the biopsy specimen. In this case, the molecular pathology test, including next-generation sequencing, may be helpful. If pathologist can identify some mutation in cells or cystic fluid, differential diagnosis of cystic neoplasm may be possible. These are KRAS and GNAS, VHL, and CTNNB1 mutation in mucinous cystic neoplasm, intraductal papillary-mucinous neoplasm, serous cystic neoplasm, and solid pseudopapillary neoplasm, respectively. The next-generation sequencing is an emerging molecular test that can detect multiple biomarkers for diagnosis, including pancreas cystic neoplasm. It has been reported that next-generation sequencing test can be applied for differential diagnosis of pancreas cystic neoplasm. However, these molecular pathology tests were not all-around; it needs to be properly managed with pathologist's quality control. It should be remembered that even if it goes through quality control, it may show a failure rate of around 30%. Despite the advances in molecular methods of high techniques, it should be remembered that the most important thing in pathologic diagnosis of pancreas cystic neoplasm is an endoscopist's skill and pathologist's expertise those provide adequate specimen and accurate diagnosis.


Subject(s)
Biomarkers , Biopsy , Diagnosis , Diagnosis, Differential , High-Throughput Nucleotide Sequencing , Mucins , Pancreas , Pancreatic Cyst , Pathology, Molecular , Quality Control
16.
Article in Korean | WPRIM | ID: wpr-764093

ABSTRACT

Thyroid nodules are the most common endocrine tumor. Ultrasonography and fine-needle aspiration (FNA) are currently accurate diagnostic tools for evaluating thyroid nodules. However, 10–30% of FNA specimens are cytologically indeterminate. Making an accurate diagnosis between benign and malignant nodules is important so that patients with malignant nodule receive proper treatment and patients with benign nodule can avoid unnecessary treatment. Several genetic changes such as point mutations of the BRAF or RAS and rearrangements of the RET/PTC1, RET/PTC3, PAX8/PPARY are used to adjust to indeterminate FNA. Such a mutational analysis has an excellent positive predictive value (PPV), but there is a weakness in the low negative predictive value (NPV). Gene-expression classifier (GEC) has been found helpful in identify nodules that are benign rather than malignant. GEC has an excellent NPV, but there is a weakness of low PPV. Multiplatform mutational and miRNA test (MPT) and next-generation sequencing assay (NGS) are being studied to compensate for these weaknesses. Molecular tests appear to be a good solution for improving the accuracy of indeterminate FNA cytology specimens and support the clinical management decisions in patients with indeterminate cytologic nodules, but further prospective multicenter trials are required for validation of reported findings and need evaluation of cost-effectiveness. This paper will review recently available molecular diagnostic tools of thyroid nodule.


Subject(s)
Biopsy, Fine-Needle , Diagnosis , Humans , MicroRNAs , Multicenter Studies as Topic , Pathology, Molecular , Point Mutation , Prospective Studies , Thyroid Gland , Thyroid Neoplasms , Thyroid Nodule , Ultrasonography
17.
Article in English | WPRIM | ID: wpr-764034

ABSTRACT

Recently, the importance of on-site detection of pathogens has drawn attention in the field of molecular diagnostics. Unlike in a laboratory environment, on-site detection of pathogens is performed under limited resources. In this study, we tried to optimize the experimental conditions for on-site detection of pathogens using a combination of ultra-fast convection polymerase chain reaction (cPCR), which does not require regular electricity, and nucleic acid lateral flow (NALF) immunoassay. Salmonella species was used as the model pathogen. DNA was amplified within 21 minutes (equivalent to 30 cycles of polymerase chain reaction) using ultra-fast cPCR, and the amplified DNA was detected within approximately 5 minutes using NALF immunoassay with nucleic acid detection (NAD) cassettes. In order to avoid false-positive results with NAD cassettes, we reduced the primer concentration or ultra-fast cPCR run time. For singleplex ultra-fast cPCR, the primer concentration needed to be lowered to 3 µM or the run time needed to be reduced to 14 minutes. For duplex ultra-fast cPCR, 2 µM of each primer set needed to be used or the run time needed to be reduced to 14 minutes. Under the conditions optimized in this study, the combination of ultra-fast cPCR and NALF immunoassay can be applied to on-site detection of pathogens. The combination can be easily applied to the detection of oral pathogens.


Subject(s)
Convection , DNA , Electricity , Immunoassay , NAD , Pathology, Molecular , Polymerase Chain Reaction , Salmonella
18.
Article in English | WPRIM | ID: wpr-760491

ABSTRACT

BACKGROUND: Extraction of cell-free DNA (cfDNA) is a key step for determining the quality of cfDNA-related molecular diagnostics. We evaluated the effect of sample containers and sample storage conditions on cfDNA extraction. METHODS: The cfDNA extraction using the MagMAX Cell-Free DNA Isolation Kit from five healthy controls and five lung cancer patients was evaluated according to the type of sample container and storage conditions: K2-EDTA container, <1, 6, 24, and 48 hr storage at 4℃ after immediate plasma separation; and Cell-Free DNA BCT container, <1, 3, 7, and 14 days stored at room temperature. Mutation analysis of EGFR exons 18–21 was performed. To assess the effect of a delay in centrifugation, EDTA whole blood samples from five healthy individuals were stored at 4℃ for 6, 12, and 24 hr before plasma separation. RESULTS: There was no significant difference in the amount and nucleic acid size of cfDNA in both controls and patients with cancer when EDTA plasma was stored at 4℃ up to 48 hr. The amount and size of cfDNA in the BCT container were not different up to 7 days; however, the 14-day sample showed an increase in cfDNA concentration due to genomic DNA contamination. EGFR mutations were detected on EDTA containers up to 48 hr and with BCT containers up to 14 days. When EDTA whole blood was stored at 4℃ and plasma separation was delayed, the cfDNA concentration increased from 24 hr. CONCLUSIONS: The cfDNA extraction was affected by the sample containers and storage conditions.


Subject(s)
Biopsy , Centrifugation , DNA Contamination , DNA , Edetic Acid , Exons , Humans , Lung Neoplasms , Pathology, Molecular , Plasma
19.
Article in English | WPRIM | ID: wpr-761735

ABSTRACT

In recent years, the taeniasis has been rarely reported in the Republic of Korea (Korea). But in this study, we intend to report 4 taeniasis cases caused by Taenia saginata during a 5-month period (February to June 2018) at a unversity hospital in Gwangju, Korea. Worm samples (proglottids) discharged from all cases were identified by phenotypic and molecular diagnostics. Mitochondrial cytochrome c oxidase subunit I sequences showed 99.4–99.9% identity with T. saginata but, differed by 4% from T. asiatica and by 7% from T. multiceps, respectively. We found that tapeworms in 2 cases (Cases 2 and 3) yielded exactly the same sequences between them, which differed from those in Cases 1 and 4, suggesting intra-species variation in tapeworms. These taeniasis cases by T. saginata infection in this study, which occurred within a limited time period and region, suggest the possibility of a mini-outbreak. This study highlights the need for further epidemiological investigation of potentially overlooked cases of T. saginata infection in Korea.


Subject(s)
Cestoda , Electron Transport Complex IV , Korea , Pathology, Molecular , Republic of Korea , Taenia saginata , Taeniasis
20.
Rio de Janeiro; s.n; 2019. 65 p. ilus.
Thesis in Portuguese | LILACS | ID: biblio-1248386

ABSTRACT

Gastrópodes continentais atuam como hospedeiros intermediários de nematódeos de interesse para a saúde humana e animal, como Angiostrongylus spp. e outros pertencentes à superfamília Metastrongyloidea. O procedimento de digestão artificial é comumente utilizado para recuperar larvas de nematódeos em moluscos. Entretanto, essas formas imaturas não apresentam desenvolvidos os caracteres morfológicos necessários à sua identificação taxonômica. Como o produto da digestão artificial é ácido e contém restos de tecido do molusco, também surgem dificuldades para realizar o diagnóstico molecular destas larvas. Estudos pilotos mostraram ineficiência no diagnóstico molecular devido a variações na qualidade da amostra, baixa concentração de DNA e baixa eficiência nas Reações em Cadeia da Polimerase (PCR). O objetivo deste trabalho foi aprimorar o diagnóstico molecular de larvas de Angiostrongylus spp. obtidas de moluscos. Assim, inicialmente, foi padronizado um método de preparo das larvas (triagem das larvas com micropipeta, somado ao ajuste do pH com PBS 1X ). A partir destas amostras padronizadas quanto ao método de preparo, foram testados métodos de extração de DNA e diferentes iniciadores para a região do Citocromo c Oxidase subunidade I (COI). Foram utilizadas larvas L3 de Angiostrongylus cantonensis, A. costaricensis e A. vasorum provenientes de ciclos mantidos em laboratório.


Amostras do campo de Aelurostrongylus abstrusus também foram utilizadas para comparação. A importância do preparo da amostra foi testada através de reações de PCR realizadas em condições idênticas, com amostras padronizadas e não padronizadas quanto ao preparo. Quanto aos testes de extração, após a realização de pilotos com diferentes protocolos, foram selecionados para testes comparativos, até a etapa de sequenciamento, dois protocolos com choque térmico e um utilizando kit comercial. Estes foram avaliados quanto à quantidade de DNA, eficiência da amplificação, tempo de realização e qualidade da sequência. Em uma terceira etapa foram testados três diferentes iniciadores para o COI. Os resultados demonstraram que após uma lavagem com PBS o produto da digestão artificial torna-se neutro (pH 7,29 ± 0,13) e que estas amostras, devidamente triadas, apresentam taxa de amplificação 60% maior. Este passo mostrou-se essencial para o sucesso das demais etapas. As amostras que foram submetidas à extração de DNA com os três protocolos selecionados resultaram em um produto que permitiu a amplificação do fragmento do COI nas reações de PCR, apesar das baixas concentrações de DNA, tendo sido possível também o diagnóstico molecular, por sequenciamento de Sanger, de 100% dessas amostras.


Os protocolos com choque térmico apresentaram menor tempo de execução, custo e geração de resíduo químico. Os três iniciadores testados amplificaram com eficiência a região-alvo, viabilizando a identificação de Angiostrongylus spp. por sequenciamento. Os diferentes métodos de extração e iniciadores testados representam diferentes possibilidades para o diagnóstico de larvas de Angiostrongylus spp. e seu uso deverá levar em conta os recursos disponíveis (tempo e dinheiro) e o objetivo do estudo (diagnóstico ou pesquisa). A disponibilização de um protocolo eficiente e a possibilidade de redução dos custos no diagnóstico dos nematódeos associados a moluscos contribuem para incentivar as pesquisas na área e viabilizar estudos de vigilância epidemiológica e controle das angiostrongilíases. (AU)


Subject(s)
Animals , Gastropoda , Pathology, Molecular , DNA Barcoding, Taxonomic , Angiostrongylus , Nematoda
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