scholarly journals PERBANDINGAN DETEKSI PLASMODIUM SPP ANTARA METODE IMMUNOCHROMATOGRAPHIC ASSAY DENGAN METODE POLYMERASE CHAIN REACTION

2014 ◽  
Vol 2 (1) ◽  
Author(s):  
Johanes Nyoman D. Widiswara Mawan

Abstract: Malaria is caused by protozoa of the genus Plasmodium remains a health problem in the world, especially in tropical countries and subtropical. Incidence of malaria from the World Health Organization (WHO) shows that in 2010 as many as 219 million cases of clinical malaria episodes show and 660,000 of them died. Therefore we need a means of early diagnosis has a sensitivity and specificity are good. This study compared the sensitivity and specificity of detection of Plasmodium spp using Immunochromatographic Assay method commonly known as rapid inspection test and Polymerase Chain Reaction (PCR). This study is a diagnostic test with a sample of 30 people who were taken with random sampling method in malaria patients who come to Budi Mulia Hospital since September 2013 - November 2013. The sample is a blood specimen taken at the brachial vein previously given informed consent in patients with the triad of symptoms of malaria in the area of ​​Bitung, Manado. From the blood samples examined by PCR. The results of the rapid tests and PCR in the detection of Plasmodium spp diagnostic test is then performed to determine the level of sensitivity and specificity. Result: The level of sensitivity of rapid tests in general by 89,2%, specificity of 100%, a positive predictive value of 100% and a negative predictive value of 40%. Conclusions: The sensitivity is moderate but has high specificity. Keywords:   Immunochromatographic Assay, Polymerase Chain Reaction (PCR), rapid tests, sensitivity, specificity  Abstrak: Malaria yang disebabkan oleh protozoa dari genus Plasmodium masih menjadi masalah kesehatan di dunia terutama di negara- negara tropis dan subtropis. Kejadian malaria dari World Health Organization (WHO) menunjukan bahwa pada 2010 sebanyak 219 juta kasus menunjukan episode klinik malaria dan 660.000 diantaranya meninggal dunia. Oleh karena itu diperlukan suatu alat diagnosa dini yang memiliki sensitivitas dan spesifisitas yang yang baik. Penelitian ini membandingkan tingkat sensitivitas dan spesifisitas deteksi Plasmodium spp dengan menggunakan metode Immunochromatographic Assay yang biasa dikenal dengan pemeriksaan rapid tes dan Polymerase Chain Reaction (PCR). Penelitian ini merupakan penelitian uji diagnostik dengan sampel sejumlah 30 orang yang diambil secara random sampling pada pasien malaria yang datang ke RSU Budi Mulia sejak bulan September 2013 - November 2013. Sampel adalah spesimen darah yang diambil pada vena brachialis yang sebelumnya telah diberikan inform consent pada pasien dengan gejala trias malaria di daerah Bitung, Manado. Dari sampel darah tersebut dilakukan pemeriksaan dengan PCR. Hasil dari rapid tes dengan metode Immunochromatographic dan PCR dalam mendeteksi Plasmodium spp selanjutnya dilakukan uji diagnostik untuk mengetahui tingkat sensitivitas dan spesifisitasnya. Hasil : Tingkat sensitivitas rapid tes secara umum sebesar 89,2%, spesifisitas sebesar 100%, nilai duga positif sebesar 100% dan nilai duga negatif sebesar 40%. Simpulan: Nilai sensitivitas yang sedang tetapi  memiliki  nilai spesifisitas  yang tinggi. Kata Kunci : Immunochromatographic Assay, Polymerase Chain Reaction (PCR), rapid tes, sensitivitas, spesifisitas

2021 ◽  
pp. 35-39
Author(s):  
Hanna Sahhar ◽  
Karly Derwitz ◽  
Erica Rubin

Since the declaration of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic in March 2020 by the World Health Organization (WHO), there has been an emergence of a new syndrome termed multisystem inflammatory syndrome in children (MIS-C) associated with COVID-19. MIS-C is defined by the presence of fever, systemic inflammation and multiorgan dysfunction in association with SARS-CoV-2 infection or COVID-19 exposure. Knowledge of this syndrome’s presentation and pathophysiology is constantly evolving as more cases are reported in the literature. This case identifies a 3-month-old patient who tested negative for SARS-CoV-2 antigen, reverse transcriptase polymerase chain reaction (RT-PCR) and antibodies but qualified for MIS-C diagnosis. To the best of our knowledge and through extensive research at the time of diagnosing and reporting this condition to the healthcare authorities, we report the youngest pediatric patient with MIS-C diagnosis. We document this case to contribute to further understanding the variable manifestations of MIS-C and the importance of early diagnosis and treatment with intravenous immunoglobulin (IVIG).


2012 ◽  
Vol 70 (9) ◽  
pp. 686-690 ◽  
Author(s):  
Carla Maria Sena Andrade Slater ◽  
Luiz Claudio Pereira Ribeiro ◽  
Marzia Puccioni-Sohler

The World Health Organization recommends the use of Osame's criterion (1990) for the diagnosis of HTLV-I-associated myelopathy (HAM/TSP). In 2006, a group of neurologists developed a Brazilian criterion that can diagnose HAM/TSP from its onset. OBJECTIVE: It was to test the agreement between both criteria. METHODS: The study included evaluation of clinical and laboratory findings of 35 patients. The ELISA, Western blot and/or polymerase chain reaction was used to search for anti-HTLV-I antibodies. The analysis of agreement was based on the calculation of Kappa. RESULTS: Concordance of 100% (Kappa=1) occurred in cases of "defined" HAM/TSP, but not in patients with "probable" diagnosis. CONCLUSION: The Brazilian criteria was as effective as Osame's criteria for the diagnosis of "defined" HAM/TSP. However, both require more specific biological markers in cerebrospinal fluid for the laboratory diagnosis of probable cases.


2021 ◽  
Vol 29 (3) ◽  
pp. 299-306
Author(s):  
Simona Roxana Gheorghe ◽  
Cătălin Marian ◽  
Ligia Gabriela Tătăranu ◽  
Anica Dricu ◽  
Cees Vermeer ◽  
...  

Abstract Meningiomas are classified by the World Health Organization (WHO) in three grades, based on morphological features. Independent of this grading, the presence of calcification in meningiomas influences their growth rate. The messenger RNA of matrix Gla protein (MGP), an extra-hepatic protein with different conformations involved in the homeostasis of ectopic calcification has been found in meningiomas and was shown to be regulated in breast cancer by miR-155-5p, a specific micro RNA. Therefore, we investigated the expression of miR-155-5p and its relationship with local MGP conformations in different grade meningiomas. According to the WHO classification, our 41 samples of meningiomas were stratified in groups WHO I and WHO II. Using real time polymerase chain reaction, we observed a higher miR-155-5p expression in group WHO I versus group WHO II [with a fold change (FC) of 3.83, p=0.027)]. Moreover, the expression of miR-155-5p was higher in calcified tumors compared to non-calcified tumors in all samples (FC=3.01, p=0.047) and in group WHO I (FC=3.65, p=0.048). Utilizing immunohistochemistry, we determined the concurrent presence of all MGP conformations in calcified meningiomas. This study was the first to establish higher miR-155-5p expression in grade WHO I and calcified meningiomas, which could improve molecular classification and targeted therapy and also the presence of all MGP conformations in calcified meningiomas, confirming the existence of an anti-calcification mechanism in meningiomas..


Author(s):  
Cristina Bragança ◽  
Inês Gonçalves ◽  
Luísa Guerreiro ◽  
Maria Janeiro

AbstractTuberculosis is an infectious disease caused by Mycobacterium tuberculosis. According to data from the World Health Organization, this disease remains one of the leading causes of death worldwide. Although it most commonly affects the lungs, tuberculosis can compromise any organ. The present study reports a rare case of vulvar tuberculosis in a postmenopausal woman with a history of asymptomatic pulmonary and pleural tuberculosis, with no prior documented contact with the bacillus. Diagnosis was based on vulvar lesion biopsies, with histological findings suggestive of infection and isolation of M. tuberculosis by microbiological culture and polymerase chain reaction (PCR) essays. The lesions reverted to normal after tuberculostatic therapy.


2020 ◽  
Vol 10 (2) ◽  
pp. 90-92
Author(s):  
Rano Mal Piryani ◽  
Suneel Piryani ◽  
Shomeeta Piryani ◽  
Ganesh Dangal ◽  
Muzaherul Huq ◽  
...  

COVID-19 is mainly transmitted through droplet infection and spread very fast compared to SARS-CoV and MERS-CoV. For the countries, it is important to know at what stage the COVID-19 epidem­ic is? So, as to take appropriate steps to contain the epidemic. This will only be known by testing the suspects and contacts of confirmed cases. If there is poor testing, then most of the infected people may remain undetected, however they could spread the virus to hundreds of other people and potential contacts, which could not be known and quarantined in time continuing the spread. If there is quality assured, highly sensitive and specific testing along with adequate isolation and quarantine, then the spread will be limited. There are two types of tests available for COVID-19: the tests directly detecting the viral ribonucleic acid (RNA) collected in nasopharyngeal or throat swabs, and tests detecting antibodies from the blood sample. At this point in time, the polymerase chain reaction (PCR) tests are used for confirmation of the disease while antibodies tests may provide information regarding the prevalence of infection. World Health Organization advices the countries to increase the testing and get to know the level of epidemic and act accordingly for containment of infection.


2021 ◽  
Vol 19 (2) ◽  
pp. 147-151
Author(s):  
M. Kunchev ◽  
V. Belcheva ◽  
E. Grigorov

Q fever, which is caused by Coxiella burnetii, a small, pleomorphic intracellular bacterium, is the most widespread zoonosis in the world. The chronic form of the disease can lead to disability and death. Rapid diagnosis of Q fever is needed in order that effective treatment can be initiated. The conventional retrospective diagnosis of Q fever, based on serology, is useless for the treatment of afflicted patients. Thus, molecular methods have been created to close the diagnostic gap between the onset of the disease and the presence of specific antibodies in serum. A polymerase chain reaction is a suitable and reliable method with high sensitivity and specificity, but it requires expensive equipment and post-amplification protocol. Loop-mediated isothermal amplification (LAMP) is an isothermal technique, conducted at constant temperature that can amplify a negligible amount of DNA to more than 109 copies within one hour, using special primers and polymerase. We have tested the sensitivity and specificity of LAMP in the detection of C. burnetii. The mean positive rate of LAMP and polymerase chain reaction in patients was 100% and 74%, respectively. LAMP reacted negatively with non-C. burnetii pathogens and non-infected blood samples. We conclude that LAMP is a sensitive and specific technique for the detection of C. burnetii and has advantages over serological methods and PCR that make it attractive for diagnosing Q fever in countries around the world.


2019 ◽  
Vol 18 ◽  
pp. 153303381982839 ◽  
Author(s):  
Moritz Perrech ◽  
Lena Dreher ◽  
Gabriele Röhn ◽  
Pantelis Stavrinou ◽  
Boris Krischek ◽  
...  

To date, diagnosis of IDH1 mutation is based on DNA sequencing and immunohistochemistry, methods limited in terms of sensitivity and ease of use. Recently, the diagnosis of IDH1 mutation by real-time polymerase chain reaction was introduced as an alternative method. In this study, real-time polymerase chain reaction was validated as a tool for detection of IDH1 mutation, and expression levels were analyzed for correlation with course of the disease. A total of 113 tumor samples were obtained intraoperatively from 84 patients with glioma having a diagnosis of diffuse glioma (World Health Organization II), anaplastic glioma (World Health Organization III), secondary glioblastoma ± chemotherapy, primary glioblastoma ± chemotherapy (World Health Organization IV). Tumor samples were snap frozen and processed for sectioning and RNA and protein isolation. Presence of IDH1 mutation was determined by DNA sequencing. Hereafter, quantitative expression of IDH1 messenger RNA was assessed using real-time polymerase chain reaction with specific primers for IDH1 mutation and –wt; protein expression was verified by Western Blot analysis and immunohistochemistry. Additionally, 19 samples of low-grade glioma and their consecutive high-grade glioma were analyzed at different time points of the disease. IDH1 mutation was identified in 63% of samples by DNA sequencing. In correlation with the real-time polymerase chain reaction results, a cutoff value was determined. Above this threshold, sensitivity and specificity of real-time polymerase chain reaction in detecting IDH1 mutation were 98% and 94%, respectively. Quantitative analysis revealed that IDH1 mutation expression is upregulated in secondary glioblastoma (mean ± standard error of mean: 3.52 ± 0.55) compared to lower grade glioma (II = 1.54 ± 0.22; III = 1.67 ± 0.23). In contrast, IDH1 wt expression is upregulated in all glioma grades (concentration >0.1) compared to control brain tissue (0.007 ± 0.0016). Western Blot analysis showed a high concordance to both sequencing and real-time polymerase chain reaction results in qualitative analysis of IDH1 mutation status (specificity 100% and sensitivity 100%). Moreover, semiquantitative protein expression analysis also showed higher expression levels of mutated IDH1 in secondary glioblastoma. In our study, real-time polymerase chain reaction and Western Blot analysis were found to be highly efficient methods in detecting IDH1 mutation in glioma samples. As cost-effective and time-saving methods, real-time polymerase chain reaction and Western Blot analysis may therefore play an important role in IDH1 mutation analysis in the future. IDH1 mutation expression level was found to correlate with the course of disease to a certain extent. Yet, clinical factors as recurrent disease or prior radiochemotherapy did not alter IDH1 mutation expression level.


Blood ◽  
2010 ◽  
Vol 116 (22) ◽  
pp. e111-e117 ◽  
Author(s):  
Helen E. White ◽  
Paul Matejtschuk ◽  
Peter Rigsby ◽  
Jean Gabert ◽  
Feng Lin ◽  
...  

Serial quantitation of BCR-ABL mRNA levels is an important indicator of therapeutic response for patients with chronic myelogenous leukemia and Philadelphia chromosome–positive acute lymphoblastic leukemia, but there is substantial variation in the real-time quantitative polymerase chain reaction methodologies used by different testing laboratories. To help improve the comparability of results between centers we sought to develop accredited reference reagents that are directly linked to the BCR-ABL international scale. After assessment of candidate cell lines, a reference material panel comprising 4 different dilution levels of freeze-dried preparations of K562 cells diluted in HL60 cells was prepared. After performance evaluation, the materials were assigned fixed percent BCR-ABL/control gene values according to the International Scale. A recommendation that the 4 materials be established as the first World Health Organization International Genetic Reference Panel for quantitation of BCR-ABL translocation by real-time quantitative polymerase chain reaction was approved by the Expert Committee on Biological Standardization of the World Health Organization in November 2009. We consider that the development of these reagents is a significant milestone in the standardization of this clinically important test, but because they are a limited resource we suggest that their availability is restricted to manufacturers of secondary reference materials.


2021 ◽  
Author(s):  
WURI RATNA HIDAYANI

Kasus pneumonia misterius yang belum diketahui etiologinya dilaporkan pada akhir Desember 2019 di Tiongkok. Terjadi lonjakan kasus dalam waktu relative singkat pasien pneumonia berjumlah 44 pasien dan terus meningkat menjadi ribuan kasus. Kasus ini kemudian diidentifikasikan ke dalam kasus Novel Corona Virus Diseases 19 (COVID 19) (Perhimpunan Dokter Paru Indonesia, 2020). Menurut World Health Organization (WHO) menyatakan bahwa hasil diagnosis tes Polymerase Chain Reaction (PCR) negative dengan tanda klinis infeksi COVID 19 di Kazakhstan dikelompokkan ke dalam perhitungan total COVID 19, pernyataan ini disiarkan sejak tanggal 1 Agustus 2020 dengan laporan adanya kasus pneumonia dengan gejala klinis COVID 19 sebanyak 13.121 kasus dengan kematian sebanyak 152 kematian sehingga Case Fatality Rate (CFR) sebesar 1,16% (Xinshua, 2020). Pada Tahun 2015 di dunia kasus pneumonia mencapai 920.000 jiwa setiap tahunnya dengan arti bahwa ada 2 balita meninggal setiap menitnya. Menurut WHO (2017) menyatakan bahwa terdapat 25.481 kematian karena pernafasan akut atau 17% dari seluruh kematian dunia dan Indonesia merupakan peringkat 7 dunia pada kasus pneumonia (Newswire, 2019).


2020 ◽  
Vol 23 (7) ◽  
pp. 498-502 ◽  
Author(s):  
Huda Fatima Rajani ◽  
Fatima Ahmed Alshaikh ◽  
Amir Anushiravani

A recent outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has affected more than 1100000 (April 5, 2020) individuals worldwide and is spreading rapidly. The virus is reported to be derived from bats and the infection was first reported in China. Similar to the severe acute respiratory syndrome and the Middle East respiratory syndrome coronaviruses, it is responsible for respiratory tract infection. Real time polymerase chain reaction and radiography are the two main diagnostic methods. Guidelines from the Center for Disease Control and Prevention and the World Health Organization (WHO) should be followed for diagnostic and precautionary measures. Treatment of the infection is still not available; however, antivirals are under clinical trials.


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