Diagnosis of intestinal microsporidiosis in pediatric oncology patients in Egypt using modified acid fast trichrome staining versus PCR

2011 ◽  
Vol 56 (4) ◽  
Author(s):  
Hadir El-Mahallawy ◽  
Mayssa Zaki ◽  
Maha El-Arousy ◽  
Lobna Shalabi ◽  
Tarek Mansour
Keyword(s):  
Clinical Laboratory ◽  
Chronic Diarrhea ◽  
Staining Technique ◽  
Negative Control ◽  
Molecular Techniques ◽  
Specific Staining ◽  
Patients With Cancer ◽  
Pediatric Cancer Patients ◽  
Stool Samples ◽  
Intestinal Microsporidiosis

AbstractMicrosporidia are intracellular opportunistic parasites that cause chronic diarrhea in AIDS patients. Little is known about the prevalence of these pathogens in pediatric cases with cancer and diarrhea. Unidentified causes of chronic diarrhea were previously encountered in pediatric cancer patients at the National Cancer Institute in Egypt. Therefore, this study tried to search for the contribution of microsporidia as a causative agent of diarrhea in this population using acid-fast trichrome stain as a specific staining and the PCR in order to evaluate the staining technique in clinical diagnosis of microsporidia. Between January 2008 and June 2009, 271 diarrheic samples from pediatric patients with cancer were studied. Microsporidia were confirmed in 13 (4.8%) cases by both PCR and staining, and additional 2 samples were positive only by staining. As a negative control, stool samples from 60 diarrheic children without malignant cancer and no microsporidia detection were examined by these two methods. So, it can be concluded that adding a diagnostic test for microsporidia to the clinical laboratory work in hospitals concerned with cancer is essential. Acid fast trichrome staining technique as being nearly efficacious as the PCR, but simpler and less expensive, can replace the molecular techniques for the diagnosis of microsporidia.

scholarly journals Microsporidiosis in a Brazilian University Hospital: case report

2006 ◽  
Vol 48 (6) ◽  
pp. 351-352 ◽  
Author(s):  
Elenice Messias do Nascimento Gonçalves ◽  
Iaiko Horroiva Uemura ◽  
Magali Orban ◽  
Vera Lúcia Pagliusi Castilho ◽  
Carlos Eduardo Pereira Corbett
Keyword(s):  
Case Report ◽  
Clinical Laboratory ◽  
Chronic Diarrhea ◽  
Staining Technique ◽  
University Hospital ◽  
Etiological Diagnosis ◽  
Cd4 Cell Count ◽  
Stool Specimens ◽  
Cd4 Cell ◽  
Concentration Methods

This is the report on a patient with chronic diarrhea caused by microsporidia. He is married, infected with HIV and has low CD4 cell count. The diagnosis was established through stool parasite search using concentration methods and Gram - chromotrope staining technique. Ileum biopsy was also performed in this case. The etiological diagnosis may be established in a clinical laboratory, by chromotrope staining technique in routine microscopic examination of stool specimens.

Quantitative alkaline phosphatase isoenzyme determination by electrophoresis on cellulose acetate membranes.

1977 ◽  
Vol 23 (1) ◽  
pp. 28-34 ◽  
Author(s):  
W H Siede ◽  
U B Seiffert
Keyword(s):  
Alkaline Phosphatase ◽  
Cellulose Acetate ◽  
Clinical Laboratory ◽  
Kinetic Assay ◽  
Specific Staining ◽  
Cellulose Acetate Membranes ◽  
Alkaline Phosphatase Isoenzymes ◽  
Elution Method ◽  
Routine Clinical Laboratory ◽  
Alkaline Phosphatase Isoenzyme

Abstract We present a new method for quantitative determination of alkaline phosphatase isoenzymes. This method consists of electrophoretic separation on cellulose acetate membranes, special fixation technique to avoid elution and diffusion of enzyme protein during incubation, specific staining, and quantitative evaluation by densitometric measurement. We highly recommend the precedure for routine clinical laboratory use. In all normal individuals we observe two isoenzymes of hepatic origin and one isoenzyme each of osseous, intestinal, and biliary origin. Quantitative normal values are presented. Precision of the method is calculated, the CV being less than 10%. The exactness of densitometric quantification is proved by comparison with kinetic assay of alkaline phosphatase isoenzymes by use of an elution method. Clinical implications of alkaline phosphatase isoenzymograms are reported and discussed in detail.

scholarly journals Enteroparasitic occurrence in fecal samples analyzed at the University of Western São Paulo-UNOESTE clinical laboratory, Presidente Prudente, São Paulo State, Brazil

2004 ◽  
Vol 46 (5) ◽  
pp. 243-248 ◽  
Author(s):  
Nair Toshiko Tashima ◽  
Maria Jacira Silva Simões
Keyword(s):  
Giardia Lamblia ◽  
Clinical Laboratory ◽  
Clinical Signs ◽  
Data Bank ◽  
Sao Paulo ◽  
São Paulo ◽  
Bank Structure ◽  
Stool Samples ◽  
Medical Attendance ◽  
The University

This study aims to analyze the enteroparasitic occurrence in children from 0 to 12 years old consulted at the University of western São Paulo Clinical Laboratory, Presidente Prudente, SP, Brazil, in relation to the socioeconomic profile of the attended children. Stool samples were examined and a questionnaire was applied with the objective of knowing the patient's age, sex, medical attendance, characteristic of the habitation, provisioning of water, dejection and domestic waste fates, use of footwear and clinical signs. The software EPI INFO 6 (Version 6.04b) was used for the elaboration of the data bank structure and analysis after previous data codification. Among 1,000 children analyzed, as many as 21.3% presented some kind of parasite. The most frequent protozoan was Giardia lamblia (7.3%) followed by Entamoeba coli (3.9%). The most frequent helminth was Enterobius vermicularis (1.9%) followed by Hymenolepis nana (0.5%). The most frequent protozoan association was Giardia lamblia / Entamoeba coli (0.9%).

A simple silver-staining technique for detecting Bence Jones proteins in unconcentrated urine.

1987 ◽  
Vol 33 (4) ◽  
pp. 561-563 ◽  
Author(s):  
J M Sheat ◽  
M A Lorier
Keyword(s):  
Silver Staining ◽  
Clinical Laboratory ◽  
Staining Technique ◽  
Silver Stain ◽  
Concentration Step ◽  
Silver Staining Technique ◽  
Bence Jones Proteins

Abstract Detection of Bence Jones proteins in urine usually involves a concentration step, followed by electrophoresis and, if necessary, immunofixation. The time-consuming and expensive concentration step can be eliminated by use of the silver-stain technique described here. This procedure, routinely used for staining unconcentrated urine, is inexpensive, sensitive, and easily performed in a clinical laboratory. Bence Jones proteins can be detected in concentrations as low as 5 mg/L.

scholarly journals A Case of Persistent Diarrhea in a Man with the Molecular Detection of Various Campylobacter species and the First Isolation of candidatus Campylobacter infans

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1003
Author(s):  
Jacky Flipse ◽  
Birgitta Duim ◽  
Janny A. Wallinga ◽  
Laetitia R. H. de Wijkerslooth ◽  
Linda van der Graaf-van Bloois ◽  
...  
Keyword(s):  
16S Rrna ◽  
Clinical Condition ◽  
Molecular Detection ◽  
Chronic Diarrhea ◽  
Immunocompromised Patient ◽  
Continuous Exposure ◽  
Persistent Diarrhea ◽  
Colon Biopsy ◽  
Stool Samples ◽  
Campylobacter Species

A man with a well-controlled HIV infection, previously diagnosed with lymphogranuloma venereum and treated for Hodgkin’s lymphoma, was suffering from chronic diarrhea. He travelled to Indonesia in the month prior to the start of complaints. Over a 15-month period, sequences related to Campylobactertroglodytis/upsaliensis, C. pinnepediorum/mucosalis/concisus and C. hominis were detected by 16S rRNA qPCR-based assays in various stool samples and in a colon biopsy. Culture revealed the first isolation of “candidatus Campylobacter infans”, a species identified recently by molecular methods only. The patient was treated with azithromycin, ciprofloxacin and tetracycline. To identify potential continuous exposure of the patient to Campylobacter, stool samples of the partner and the cat of the patient were analyzed and C. pinnepediorum/mucosalis/concisus and C. helveticus, respectively, were detected. The diversity in detected species in this immunocompromised patient with a lack of repeatedly consistent findings resulted in the conclusion that not any of the Campylobacter species was the primary cause of the clinical condition. This study shows the challenges in detection and interpretation of diagnostic results regarding Campylobacter.

scholarly journals Integration of Germline Pharmacogenetics Into a Tumor Sequencing Program

2018 ◽  
pp. 1-15 ◽  
Author(s):  
Daniel L. Hertz ◽  
Andrew Glatz ◽  
Amy L. Pasternak ◽  
Robert J. Lonigro ◽  
Pankaj Vats ◽  
...  
Keyword(s):  
Genetic Information ◽  
Clinical Laboratory ◽  
Treatment Decisions ◽  
Added Value ◽  
Treatment Recommendation ◽  
Evidence Based ◽  
Patients With Cancer ◽  
Retrospective Assessment ◽  
Tumor Sequencing ◽  
Evidence Based Guidelines

Purpose Evidence-based guidelines inform treatment decisions for patients for whom germline genetic information is available. Our real-time tumor sequencing program, which makes precision treatment decisions for patients with cancer, produces matched germline information, providing a unique opportunity to efficiently implement pharmacogenetics and benefit patients. Methods The germline genetic database from the Michigan Oncology Sequencing (MI-Oncoseq) program was searched for 21 clinically actionable polymorphisms in five cancer-relevant genes: TPMT, DPYD, CYP2C19, CYP3A5, and UGT1A1. Residual germline DNA was sent to an external Clinical Laboratory Improvement Amendments–approved laboratory for confirmatory genotyping. The medical records of MI-Oncoseq patients with actionable phenotypes were searched for receipt of relevant drugs and to determine whether having genetic information at the time of treatment would have led to a treatment recommendation. Results All nine variants in TPMT, DPYD, and CYP2C19 that were detected in MI-Oncoseq were confirmed by external genotyping. Genotype determinations could not be made for CYP3A5*3, UGT1A1*28, or UGT1A1*80. On the basis of retrospective assessment of 115 adult and pediatric patient records, 4.3% (n = 5) had a potentially clinically actionable phenotype for TPMT, DPYD, or CYP2C19 and received a relevant medication. After accounting for differences in adult and pediatric recommendations, three of these patients could have received a treatment recommendation at the time of prescribing. Conclusion Germline genotype determinations for TPMT, DPYD, and CYP2C19 can be used to make evidence-based treatment recommendations in MI-Oncoseq patients. Although the proportion of patients for whom recommendations can be made is small, this added value to MI-Oncoseq and patient care comes at no additional genotyping cost. Pharmacogenetic assessment should be integrated into tumor sequencing programs that genotype matched germline DNA; however, the complexity and additional cost of implementing pharmacogenetics remain challenging.

scholarly journals Evaluation of Two DNA Extraction Methods for Detection of Strongyloides stercoralis Infection

2018 ◽  
Vol 56 (4) ◽  
Author(s):  
Beatrice Barda ◽  
Rahel Wampfler ◽  
Somphou Sayasone ◽  
Khampheng Phongluxa ◽  
Syda Xayavong ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Strongyloides Stercoralis ◽  
Extraction Methods ◽  
Molecular Techniques ◽  
Diagnostic Methods ◽  
Content Type ◽  
Stool Samples ◽  
Dna Extraction Methods ◽  
Baermann Method

ABSTRACT Strongyloides stercoralis is present worldwide, but its prevalence is still uncertain, mainly due to the lack of sensitivity of diagnostic methods. Molecular techniques are under development, but a standardized protocol is still unavailable. We compared the sensitivity of real-time PCR, using two extraction protocols, with that of the Baermann technique. Samples were collected in the framework of the baseline screening of a randomized clinical trial evaluating moxidectin against S. stercoralis in Lao People's Democratic Republic. Two stool samples from each participant were processed by the Baermann method, and one subsample was processed by PCR. DNA was extracted using the QIAamp DNA stool minikit based on the standard protocol for the QIAamp DNA minikit (QIA) and using a modification of the QIA procedure (POL). Subsequently, all extracted samples were analyzed by real-time PCR. Overall, 95 samples were analyzed by the three diagnostic methods. Sixty-nine (72.6%) samples were positive according to the Baermann method, 25 (26.3%) by the QIA method, and 62 (65.3%) by the POL method. The sensitivities were 86% (95% confidence interval [CI], 76.7 to 92.9), 31.0% (95% CI, 21.3 to 42.6), and 78.0% (95% CI, 66.8 to 86.1) for the Baermann, QIA, and POL methods, respectively. The sensitivities calculated for each day of the Baermann method separately were 60% (48.4 to 70.8%) and 64% (52.2 to 74.2%) for days 1 and 2, respectively. In conclusion, the POL method revealed a good performance and was comparable to the Baermann test performed on two stool samples and superior to the Baermann method performed on one stool sample. Additional studies are needed to standardize a PCR protocol for S. stercoralis diagnosis.

Parasitological versus molecular diagnosis of strongyloidiasis in serial stool samples: how many?

2017 ◽  
Vol 92 (1) ◽  
pp. 12-16 ◽  
Author(s):  
E. Dacal ◽  
J.M. Saugar ◽  
T. Soler ◽  
J.M. Azcárate ◽  
M.S. Jiménez ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Correct Diagnosis ◽  
Molecular Techniques ◽  
Maximum Sensitivity ◽  
Molecular Diagnostic ◽  
Molecular Diagnostic Techniques ◽  
Molecular Technique ◽  
Asymptomatic Patients ◽  
Stool Samples

AbstractStrongyloidiasis is usually an asymptomatic disease in immunocompetent patients, caused by Strongyloides stercoralis. However, in immunocompromised patients it can produce a severe clinical profile. Therefore, a correct diagnosis is necessary in these cases and in those chronic asymptomatic patients. The low sensitivity of classical parasitological techniques requires the analysis of multiple serial stool samples. Molecular diagnostic techniques represent an improvement in the detection of the parasite. The objective of this study was to evaluate the minimum number of samples necessary to achieve maximum sensitivity by real-time polymerase chain reaction (PCR). A total of 116 stool samples from 39 patients were analysed by direct microscopic observation, agar culture, Harada–Mori and real-time PCR, in one, two, three and four or more consecutive samples. After two serial samples, 6 out of 39 patients were positive by parasitological and molecular techniques, while 16 of them were real-time PCR positive, and all the patients detected by parasitology were also detected by the molecular technique, reaching 100.00% sensitivity versus 83.00% when analysing a single sample. These data also reflect apparently low specificity (51.52%) and positive predictive value (PPV) (27.27 %) values, due to the high number of cases detected by real-time PCR and not by parasitological techniques. These cases were confirmed as true positives when analysing three, four or more samples from the same patient. In conclusion, the application of molecular techniques decreases the number of serial stool samples necessary to give a diagnosis with the maximum sensitivity.

scholarly journals Gum arabic establishes prebiotic functionality in healthy human volunteers in a dose-dependent manner

2008 ◽  
Vol 100 (6) ◽  
pp. 1269-1275 ◽  
Author(s):  
Wim Calame ◽  
Antje R. Weseler ◽  
Christer Viebke ◽  
Cal Flynn ◽  
André D. Siemensma
Keyword(s):  
Gum Arabic ◽  
Optimal Dose ◽  
Positive Control ◽  
Negative Control ◽  
Dose Effect ◽  
Dependent Manner ◽  
Daily Consumption ◽  
Healthy Human ◽  
Human Volunteers ◽  
Stool Samples

The present study was undertaken to determine the prebiotic efficacy of gum arabic upon consumption by man for up to 4 weeks and, if any, to establish the dose–effect relationship. Human healthy volunteers consumed various daily doses (5, 10, 20, 40 g) of gum arabic (EmulGold®) in water for up to 4 weeks. Daily consumption of water was taken as the negative control and that of 10 g inulin as the positive control. At 0, 1, 2 and 4 weeks quantification of bacterial numbers in stool samples was performed via real time-PCR techniques and questionnaires were filled in to account for potential drawbacks. The genera of Bifidobacteria and Lactobacilli were taken as potentially beneficial bacteria and those of Bacteroides,Clostridium difficileand Enterococci as potentially non-beneficial, this distinction was dependent on the issue of these numbers being or becoming out of balance in the host. Compared with the negative control the numbers of Bifidobacteria and Lactobacilli 4 weeks after consumption were significantly higher for gum arabic: the optimal dose being around 10 g. Moreover, at this dose the numbers of Bifidobacteria, Lactobacilli and Bacteroides were significantly higher for gum arabic than for inulin. No significant drawback was encountered during the study. It is concluded that gum arabic establishes prebiotic efficacy, at least as good as inulin. The optimal daily dose was found to be 10 g.

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