Efficacy of infected tissue sample brief-culture on Lowenstein-Jensen media as pre-polymerase chain reaction (Pre-PCR) to diagnosis of Mycobacterium

2012 ◽  
Vol 6 (12) ◽  
Author(s):  
Fatemeh Fallah
Keyword(s):  
Polymerase Chain Reaction ◽  
Tissue Sample ◽  
Infected Tissue ◽  
Chain Reaction ◽  
Lowenstein Jensen ◽  
Polymerase Chain

scholarly journals TRC4 Gene Based PCR Assay in Diagnosis of Tuberculous Meningitis

2017 ◽  
Vol 8 (2) ◽  
pp. 19-22
Author(s):  
Abu Naser Ibne Sattar ◽  
Sanjida Khondakar Setu ◽  
Ahmed Abu Saleh ◽  
Sharmeen Ahmed
Keyword(s):  
Polymerase Chain Reaction ◽  
Clinical Manifestation ◽  
Tuberculous Meningitis ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Early Stages ◽  
Lowenstein Jensen ◽  
Polymerase Chain ◽  
Tb Pcr ◽  
Is6110 Element

The prevalence of Tuberculous meningitis remains largely underestimated due to nonspecific clinical manifestation at early stages. A total of 20 CSF samples were studied from clinically suspected cases of tuberculous meningitis. All the samples were processed for ZN staining, TB culture on LJ media and TB-PCR using IS6110 and TRC4 primers by standard protocols. Of the total 20 CSF samples, 12 cases were diagnosed as TB meningitis. Most of the tuberculous meningitis cases were found positive by PCR using TRC4 primers (83.33%) followed by IS6110 primer (66.67%) and culture on L-J media(8.33%). None were found positive by ZN staining. TB-PCR usingTRC4 primer showed higher positivity than using IS6110 in detecting tuberculous meningitis, since some strains of MTB may lack the IS6110 element in their genome. PCR assay using TRC4 primer is superior in diagnosing tuberculous meningitis. This study was aimed to evaluate the diagnostic potentials of CSF polymerase chain reaction (PCR) by using TRC4 and IS6110 primers. Further the results were also compared with culture on Lowenstein-Jensen (LJ) media and AFB smear by Zeihl-Nelson (ZN) staining.Bangladesh J Med Microbiol 2014; 08 (02): 19-22

A Microdissection and Molecular Genotyping Assay to Confirm the Identity of Tissue Floaters in Paraffin-Embedded Tissue Blocks

2003 ◽  
Vol 127 (2) ◽  
pp. 213-217 ◽  
Author(s):  
Jennifer L. Hunt ◽  
Patricia Swalsky ◽  
E. Sasatomi ◽  
Laura Niehouse ◽  
Anke Bakker ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Tissue Sample ◽  
Small Samples ◽  
Allelic Loss ◽  
Chain Reaction ◽  
Neoplastic Tissue ◽  
Tissue Samples ◽  
Genotyping Assay ◽  
Tissue Identification ◽  
Polymerase Chain

Abstract Context.—A recurring problem in surgical pathology practice is specimen mix-up and floater contamination. While many cases can be resolved histologically, a significant number remain unclear and may have serious clinical and medicolegal implications. Objectives.—To design a microdissection and genotyping assay to identify contaminating floater tissues in paraffin-embedded tissues that is optimized for small samples, and to use the assay to resolve a series of clinical cases with floater tissues. Materials and Methods.—Twenty-one cases of possible tissue floater contamination in paraffin-embedded tissue blocks were included. Using 4 unstained, 4-μm-thick histologic sections, multiple sites were microdissected under direct visualization either by hand or by laser capture microdissection. Nonneoplastic and neoplastic tissues were sampled. Polymerase chain reaction was performed for a panel of 10 polymorphic microsatellite markers at 1p34, 3p26, 5q21, 9p21, 10q23, and 17p13. Allele size and content were analyzed semiquantitatively by fluorescent capillary electrophoresis, and the genotypes for the tissues in the paraffin-embedded tissue blocks were compared for identity. Results.—Tissue identification was successful in all cases, despite small tissue sample size and fixation effects. Comparative analysis of neoplastic tissue floaters and the presumptive source tumor was performed when possible to control for possible allelic loss or microsatellite instability. Conclusions.—Microdissection and genotyping are effective and reliable means to objectively resolve problems of possible floater contamination. Even minute tissue samples provide sufficient DNA template for polymerase chain reaction microsatellite analysis. Because of the potential clinical implications of floaters, we recommend that all suspected floaters that would change a diagnosis from benign to malignant be subjected to genotyping assay to confirm the identity of the floater tissue.

scholarly journals SARS–CoV-2 isolation from an appendix

2020 ◽  
Vol 2020 (8) ◽  
Author(s):  
Shahzaib Ahmad ◽  
Rahim Nadeem Ahmed ◽  
Poonam Jani ◽  
Mattee Ullah ◽  
Hossam Aboulgheit
Keyword(s):  
Polymerase Chain Reaction ◽  
Abdominal Pain ◽  
Respiratory Symptoms ◽  
Tissue Sample ◽  
Gastrointestinal Symptoms ◽  
Shortness Of Breath ◽  
Chain Reaction ◽  
Histological Features ◽  
First Case ◽  
Polymerase Chain

Abstract Efforts to recognize SARS–CoV-2 infection have focused on respiratory symptoms such as cough and shortness of breath. Although it is also well known that SARS–CoV-2 infection can cause gastrointestinal symptoms such as abdominal pain, nausea, vomiting and diarrhoea, there are emerging reports of SARS–CoV-2 infection causing surgical pathology. We present the first case report of SARS–CoV-2 infection directly causing acute appendicitis, first suspected due to highly atypical histological features and later confirmed as polymerase chain reaction positive appendicular tissue sample.

scholarly journals A Polymerase Chain Reaction Assay for the Detection of Xanthomonas campestris pv. musacearum in Banana

Plant Disease ◽  
2010 ◽  
Vol 94 (1) ◽  
pp. 109-114 ◽  
Author(s):  
Melanie L. Lewis Ivey ◽  
Geoffrey Tusiime ◽  
Sally A. Miller
Keyword(s):  
Polymerase Chain Reaction ◽  
Xanthomonas Campestris ◽  
Pcr Primers ◽  
Infected Tissue ◽  
Conventional Pcr ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
Whole Cells ◽  
Polymerase Chain ◽  
Banana Xanthomonas Wilt

Polymerase chain reaction (PCR) primers (BXW-1 and BXW-3) for conventional PCR were developed from conserved sequences in the hrpB operon of the hrp gene cluster from Xanthomonas campestris pv. musacearum, the causative agent of banana Xanthomonas wilt (BXW). All 50 strains of X. campestris pv. musacearum, isolated from Uganda, Rwanda, and Tanzania, produced a 214-bp amplicon when whole cells, bacterial ooze from infected tissue, and genomic DNA purified from bacterial ooze or infected tissue were used as template. The BXW primers also detected strains of X. axonopodis pv. vasculorum isolated from sugarcane and maize and strains of X. vasicola pv. holcicola isolated from sorghum. All of the strains of X. campestris pv. musacearum were clonal when compared using enterobacterial repetitive intergenic consensus PCR.

scholarly journals Culture-Negative Endocarditis Diagnosed Using 16S DNA Polymerase Chain Reaction

2012 ◽  
Vol 23 (4) ◽  
pp. 216-218 ◽  
Author(s):  
Stephen Duffett ◽  
Bayan Missaghi ◽  
Peter Daley
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Polymerase ◽  
Tissue Sample ◽  
Low Grade ◽  
Chain Reaction ◽  
Severe Aortic Regurgitation ◽  
Valve Tissue ◽  
Polymerase Chain ◽  
Dna Pcr ◽  
Culture Negative

16S DNA polymerase chain reaction (PCR) is a molecular amplification technique that can be used to identify bacterial pathogens in culture-negative endocarditis. Bacterial DNA can be isolated from surgically excised valve tissue or from blood collected in EDTA vials. Use of this technique is particularly helpful in identifying the bacterial pathogen in cases of culture-negative endocarditis. A case involving a 48-year-old man who presented with severe aortic regurgitation and a four-month prodrome of low-grade fever is reported. Blood and valve tissue cultures following valve replacement were negative. A valve tissue sample was sent for investigation with 16S DNA PCR, which successfully identifiedStreptococcus salivariusand was interpreted as the true diagnosis. A review of the literature suggests that 16S DNA PCR from valve tissue is a more sensitive diagnostic test than culture. It is also extremely specific, based on a sequence match of at least 500 base pairs.

scholarly journals Métodos convencionais e moleculares para o diagnóstico da tuberculose pulmonar: um estudo comparativo

2008 ◽  
Vol 34 (12) ◽  
pp. 1056-1062 ◽  
Author(s):  
Stella Sala Soares Lima ◽  
Wanessa Trindade Clemente ◽  
Moisés Palaci ◽  
Reinaldo Vieira Rosa ◽  
Carlos Maurício de Figueiredo Antunes ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Mycobacterium Tuberculosis ◽  
Chain Reaction ◽  
Lowenstein Jensen ◽  
Polymerase Chain

OBJETIVO: Comparar quatro métodos laboratoriais no diagnóstico de tuberculose pulmonar. MÉTODOS: Foram realizadas pesquisa direta pelas colorações de Ziehl-Neelsen e auramina, cultura para micobactérias em meio Löwenstein-Jensen (LJ) e polymerase chain reaction (PCR, reação em cadeia da polimerase) para Mycobacterium tuberculosis em 160 amostras de secreção respiratória de pacientes com suspeita de tuberculose pulmonar. As cepas isoladas foram identificadas por método radiométrico utilizando-se p-nitro-alfa-acetilamino-beta-hidroxipropiofenona (NAP) e métodos clássicos. A sensibilidade dos métodos foi comparada com o padrão ouro para o diagnóstico da tuberculose pulmonar, definido por critérios clínicos, radiológicos e microbiológicos. RESULTADOS: Dos 160 pacientes, 142 foram diagnosticados com tuberculose pulmonar de acordo com o padrão ouro. As técnicas de Ziehl-Neelsen e auramina, cultura em meio LJ e PCR apresentaram sensibilidade de 54,2%, 58,4%, 67,6% e 77,5%, respectivamente, quando comparados ao critério diagnóstico adotado. A especificidade dos quatro métodos foi de 100%. A concordância na identificação da micobactéria entre PCR e o método radiométrico utilizando NAP foi alta (96,8%). A sensibilidade da PCR foi de 50,8% nas amostras com baciloscopia negativa e de 98,8% naquelas com baciloscopia positiva. Nas amostras com resultados negativos na baciloscopia e cultura, a sensibilidade da PCR foi menor que nas com resultados positivos (25,6% e 99,0%, respectivamente). CONCLUSÕES: A PCR é método promissor no diagnóstico da tuberculose pulmonar, mesmo em amostras paucibacilares. Além disso, apresenta a vantagem da identificação simultânea e rapidez do resultado.

scholarly journals Use of brief culture on Lowenstein–Jensen media as pre-polymerase chain reaction (Pre-PCR) to detect Mycobacterium

2015 ◽  
Vol 4 ◽  
pp. 90
Author(s):  
Fatemeh Fallah ◽  
Abdollah Karimi ◽  
Gita Eslami ◽  
Bakhtiar Yousefi ◽  
Bahador Yazdanpanah ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Lowenstein Jensen ◽  
Polymerase Chain

Use of Polymerase Chain Reaction to Diagnose Tuberculous Arthritis From Joint Tissues and Synovial Fluid

2004 ◽  
Vol 128 (2) ◽  
pp. 205-209 ◽  
Author(s):  
Alexei G. Titov ◽  
Elena B. Vyshnevskaya ◽  
Sergei I. Mazurenko ◽  
Seppo Santavirta ◽  
YrjöT. Konttinen
Keyword(s):  
Polymerase Chain Reaction ◽  
Outcome Measures ◽  
Tissue Sample ◽  
Bone Destruction ◽  
Sample Collection ◽  
Chain Reaction ◽  
Disease Manifestation ◽  
Tuberculous Arthritis ◽  
Lung Tuberculosis ◽  
Polymerase Chain

Abstract Context.—Tuberculosis of the joints and bones is a significant worldwide problem, often leading to joint and bone destruction. The diagnosis of this disease manifestation is difficult. Objective.—To assess the role of conventional diagnostics compared to polymerase chain reaction applied to samples obtained at arthroscopy. Design.—This was an open observational study that was blinded to the microbiologist, histopathologist, and molecular biologist responsible for assessing the main outcome measures. Patients.—Seven patients (8 samples) with joint and bone tuberculosis and 14 patients (16 samples) with nontuberculous joint and bone disease. Intervention.—Arthroscopic examination and tissue sample collection. Main Outcome Measures.—Mycobacterium tuberculosis staining, culture, and histopathologic assessment of caseating granulomas vs polymerase chain reaction. Results.—Polymerase chain reaction was positive in all cases of true tuberculosis and falsely identified 2 samples as positive, both however, in patients who had lung tuberculosis in the past. Conclusions.—Conventional bacteriological methods for demonstration of M tuberculosis are not very sensitive and can be time-consuming. Polymerase chain reaction of arthroscopically obtained joint tissue biopsies appears promising in the early diagnosis of tuberculous arthritis.

scholarly journals ASSESSMENT OF DIAGNOSTIC TECHNIQUES OF URINARY TUBERCULOSIS

2013 ◽  
Vol 5 (1) ◽  
pp. e2013034 ◽  
Author(s):  
Khaled Ismail Ghaleb ◽  
Magdy Mohamed Afifi ◽  
Mohamed Mohamed El Gohary
Keyword(s):  
Polymerase Chain Reaction ◽  
Mycobacterium Tuberculosis ◽  
Early Diagnosis ◽  
Culture System ◽  
Chain Reaction ◽  
Lowenstein Jensen ◽  
Polymerase Chain ◽  
Urinary Tuberculosis ◽  
Dna Enzyme Immunoassay ◽  
E Mail

    Khalid Ghaleb a,* , Magdy Afifib, Mohamad El-Gohary c aDepartment of Medical Laboratories, Faculty of Applied Medical Science, King Khalid University, Bisha 551, Saudia Arabia bDepartment of Botany and Microbiology, Faculty of Science, Al-Azhar University, Assuit 71524, Egypt cDepartment of Internal Medicine, Faculty of Medicine, Al-Azhar University, Assuit, Egypt The corresponding author e-mail: [email protected] Current Tel: 00966595388496 Saudia,  00201119338055 Egypt The place of the study worked : Department of Botany and Microbiology, Faculty of Science, Al-Azhar University, Assuit 71524, Egypt, e-mail: [email protected]  Tel: 00201006554961 Abstract Early diagnosis of active tuberculosis remains an elusive challenge. In addition, one third of the world's population is latently infected with Mycobacterium tuberculosis (Mtb) and up to 10% of infected individuals develop tuberculosis (TB) in their lifetime. In this investigation, the incidence of urinary tuberculosis among renal patients was studied. Three hundreds urine samples were processed for detection of Mtb by Ziehl-Neelson (ZN) smear examination, Lowenstein Jensen (LJ) medium, radiometric BACTEC460 system as well as polymerase chain reaction (PCR) and DNA Enzyme Immunoassay (DEIA) test.  Out of 300 urine samples, 2 were positive by both  ZN smears and LJ medium with incidence rate of 0.66 %, 3 positive samples by BACTEC460 culture system with incidence of 1%. PCR assay gave more positive results than smear and culture examination (i.e. 8 positive samples with incidence  rate of 2.6%).  The specificities were 25% for both ZN smears and LJ medium, 37.5% for BACTEC460 culture system, and 100% for PCR test, while  sensitivities of all assays were 100%. Thus PCR is a rapid and sensitive method for the early diagnosis of urinary tuberculosis.   Keywords: List of abbreviations:Acid Fast Bacilli (AFB)-Base pair (bp)-DNA Enzyme Immunoassay (DEIA)  -Extrapulmonary Tuberculosis (EPTB)-International Union Against Tuberculosis  (IUAT)-Lowenstein Jensen (LJ)-Mycobacterium tuberculosis (Mtb) -Polymerase Chain Reaction (PCR)-Tuberculosis (TB)-Urinary Tuberculosis (UTB-Urogenital Tuberculosis (UGTB)-Ziehl-Neelson (ZN)  

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