Investigation of Cyclospora cayetanensis by modified acid-fast staining and nested PCR in some patient groups

2021 ◽  
Vol 0 (0) ◽  
pp. 0-0
Author(s):  
Abdurrahman EKİCİ ◽  
Ahmet Hakan ÜNLÜ ◽  
Hasan YILMAZ ◽  
Zeynep TAŞ CENGİZ ◽  
Yunus BEYHAN
Keyword(s):  
Nested Pcr ◽  
Cyclospora Cayetanensis ◽  
Patient Groups ◽  
Acid Fast Staining

scholarly journals A Molecular Tool for Rapid Detection and Traceability of Cyclospora cayetanensis in Fresh Berries and Berry Farm Soils

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 261
Author(s):  
Carolina N. Resendiz-Nava ◽  
Guadalupe E. Orozco-Mosqueda ◽  
Edmundo M. Mercado-Silva ◽  
Susana Flores-Robles ◽  
Hilda V. Silva-Rojas ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Nested Pcr ◽  
Limit Of Detection ◽  
Rrna Gene ◽  
Human Pathogen ◽  
Pcr Assay ◽  
Accurate Identification ◽  
Cyclospora Cayetanensis ◽  
Molecular Tool ◽  
Accurate Detection

Due to recent outbreaks of cyclosporiasis associated with consumption of fresh berries, producers are demanding modern microbiological tools for the rapid and accurate identification of the human pathogen Cyclospora cayetanensis in berries and environmental samples. The aim of the present work was to develop a molecular tool based on a PCR approach for the rapid and accurate detection of C. cayetanensis. A nested PCR assay was validated for the amplification of a 294 bp size region of the 18S rRNA gene from C. cayetanensis. The limit of detection for the nested PCR assay was validated using 48 berry samples spiked with ~0, 10, 100, and 1000 oocyst per gram of sample. With this assay, it was possible to detect as few as 1 oocyst per gram of berry, in a 50 g sample. Sanger DNA sequencing and phylogenetic analysis were carried out to confirm the presence of C. cayetanensis in berry (n = 17) and soil (n = 5) samples. The phylogenetic analysis revealed that the C. cayetanensis sequences obtained from Mexico clustered within a group recovered from China, Peru, Guatemala-Haiti, and Japan. The PCR protocol designed in the present study could be an important tool for the rapid and accurate detection of this human pathogen in environmental and food samples.

scholarly journals Enteric Opportunistic Infection and the Impact of Antiretroviral Therapy among HIV/AIDS Patients from Tehran, Iran

2019 ◽  
Author(s):  
Hossein MASOUMI-ASL ◽  
Khadijeh KHANALIHA ◽  
Farah BOKHARAEI-SALIM ◽  
Abdoulreza ESTEGHAMATI ◽  
Saeed KALANTARI ◽  
...  
Keyword(s):  
Antiretroviral Therapy ◽  
Nested Pcr ◽  
Staining Method ◽  
Blue Staining ◽  
Hiv Positive ◽  
Medical Sciences ◽  
Aids Patients ◽  
Acid Fast Staining ◽  
Aniline Blue Staining ◽  
Hiv Aids

Background: Opportunistic parasites have been identified as human pathogens, especially in immunodeficient patients. Microsporidian and coccidian infections cause chronic diarrhea as common clinical manifestation in HIV positive patients. In this study, the frequency of opportunistic infections, including microsporidian and coccidian infections, was evaluated in HIV/AIDS patients from Tehran and phylogenic analysis was performed for E. bieneusi isolates from these patients.  Methods: One hundred and two stool samples were collected from confirmed HIV/AIDS patients, referred to Consult Center of Behavior Diseases, West Health Center, Iran University of Medical Sciences in Tehran, Iran. The samples were transferred to Research Center of Pediatric Infectious Diseases, Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences from Jan 2016 to Dec 2016. After conventional formalin-ether concentration, aniline blue staining method and acid-fast staining technique were used for detection of microsporidian spores and Cyclospora oocysts. DNA was extracted and nested PCR was performed. Results: Two (1.96%) cases were found to be positive for intestinal microsporidia infection using aniline blue staining method and were confirmed as E. bieneusi by nested PCR. One patient was found with Cyclospora cayetanensis infection by acid-fast staining method and PCR. Giardia lamblia and Blastocystis hominis were detected as non-opportunistic parasites in 1/102 (0.98%) and 2/102 (1.96%) of the HIV positive patients, respectively. Conclusion: With respect to the use of antiretroviral therapy (ART) in HIV positive patients, we found a low frequency of infection.

scholarly journals Cyclosporiasis Among School Going Children of Kathmandu Valley

1970 ◽  
Vol 11 ◽  
pp. 193-198 ◽  
Author(s):  
Shardulendra Prasad Sherchand ◽  
Dev Raj Joshi ◽  
Nabaraj Adhikari ◽  
Krishna Prasad Pant ◽  
Ramesh Pun
Keyword(s):  
Cryptosporidium Parvum ◽  
School Children ◽  
Specific Treatment ◽  
Staining Technique ◽  
Parasitic Infections ◽  
Kathmandu Valley ◽  
Cyclospora Cayetanensis ◽  
Intestinal Parasitosis ◽  
Significant Difference ◽  
Acid Fast Staining

Intestinal parasitic infections among children possess a critical public health issue in Nepal. This study was conducted to determine cyclosporiasis among school going children of Kathmandu valley. A total of 187 collected stool samples were subjected to direct microscopy, formal-ether concentration technique, Sheather’s sucrose floatation technique and modified acid fast staining technique. The incidence of cyclosporiasis, cryptosporidiosis and other intestinal parasitosis was 24.1%, 29.4% and 51.9% respectively. The cyclosporiasis was higher in female (28.4%) without any significance (p=0.190). The cyclosporiasis was found highest in age group 5-8 (25.2%) but statistically insignificant (p=0.895). Cyclosporiasis was higher in symptomatic case (25.6%) but found statistically insignificant (p=0.469). Similarly, cyclosporiasis was higher in school children without following hygienic practice (28.0%) but without any significance (p=0.103). There was significant difference between detection of cyclospora before and after sporulation by modified acid fast staining technique (p<0.001) and highest association was seen between Cyclospora cayetanensis and Cryptosporidium parvum. The outcome of this study revealed that cyclosporiasis remain highly endemic in school children of Kathmandu valley. This suggests proper diagnosis and specific treatment is required to lower the parasitic burden. Key words: Cyclospora cayetanensis; Cryptosporidium parvum; modified acid fast staining technique DOI: 10.3126/njst.v11i0.4145Nepal Journal of Science and Technology 11 (2010) 193-198

scholarly journals Evaluation of Diagnostic Value and Epidemiological Implications of PCR for Pneumocystis carinii in Different Immunosuppressed and Immunocompetent Patient Groups

2000 ◽  
Vol 38 (4) ◽  
pp. 1461-1467 ◽  
Author(s):  
Andreas Sing ◽  
Karlheinz Trebesius ◽  
Andreas Roggenkamp ◽  
Holger Rüssmann ◽  
Karin Tybus ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Pneumocystis Carinii ◽  
Diagnostic Value ◽  
Immunocompetent Patient ◽  
Immunosuppressed Patient ◽  
Hiv Patients ◽  
Additional Advantage ◽  
Predictive Values ◽  
Patient Groups ◽  
Immunocompetent Patients

To evaluate the value of single and nested PCRs for diagnosis ofPneumocystis carinii pneumonia (PCP) in a variety of respiratorily distressed patient groups, 574 respiratory samples from 334 patients (89 human immunodeficiency virus [HIV]-positive patients, 61 transplant recipients, 66 malignancy patients, 34 otherwise immunosuppressed patients, and 84 immunocompetent patients) were prospectively examined by microscopy and single and nested PCRs. The resulting data were correlated with clinical evidence of PCP. Microscopy and single PCR of bronchoalveolar lavage (BAL) specimens from HIV patients were 100% sensitive and specific in detecting PCP, whereas nested PCR, although being 100% sensitive, reached a specificity of only 97.5%. In the three non-HIV immunosuppressed patient groups, both single and nested PCR invariably produced lower positive predictive values than microscopy. Among immunocompetent patients, the positive predictive values of both PCRs were 0%. Therefore, the diagnostic values of the PCR methods tested do not seem to offer any additional advantage compared to that of conventional microscopy for these patient groups. However, nested PCR identified a significant percentage of clinically silentP. carinii colonizations in about 17 to 20% of immunocompetent and immunosuppressed non-HIV patients.

Evaluation of an Improved U.S. Food and Drug Administration Method for the Detection of Cyclospora cayetanensis in Produce Using Real-Time PCR

2017 ◽  
Vol 80 (7) ◽  
pp. 1133-1144 ◽  
Author(s):  
Helen R. Murphy ◽  
Seulgi Lee ◽  
Alexandre J. da Silva
Keyword(s):  
Real Time ◽  
Drug Administration ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Food And Drug Administration ◽  
Fresh Produce ◽  
Pcr Assay ◽  
Detection Rates ◽  
Cyclospora Cayetanensis ◽  
The Real

ABSTRACTCyclospora cayetanensis is a protozoan parasite that causes human diarrheal disease associated with the consumption of fresh produce or water contaminated with C. cayetanensis oocysts. In the United States, foodborne outbreaks of cyclosporiasis have been linked to various types of imported fresh produce, including cilantro and raspberries. An improved method was developed for identification of C. cayetanensis in produce at the U.S. Food and Drug Administration. The method relies on a 0.1% Alconox produce wash solution for efficient recovery of oocysts, a commercial kit for DNA template preparation, and an optimized TaqMan real-time PCR assay with an internal amplification control for molecular detection of the parasite. A single laboratory validation study was performed to assess the method's performance and compare the optimized TaqMan real-time PCR assay and a reference nested PCR assay by examining 128 samples. The samples consisted of 25 g of cilantro or 50 g of raspberries seeded with 0, 5, 10, or 200 C. cayetanensis oocysts. Detection rates for cilantro seeded with 5 and 10 oocysts were 50.0 and 87.5%, respectively, with the real-time PCR assay and 43.7 and 94.8%, respectively, with the nested PCR assay. Detection rates for raspberries seeded with 5 and 10 oocysts were 25.0 and 75.0%, respectively, with the real-time PCR assay and 18.8 and 68.8%, respectively, with the nested PCR assay. All unseeded samples were negative, and all samples seeded with 200 oocysts were positive. Detection rates using the two PCR methods were statistically similar, but the real-time PCR assay is less laborious and less prone to amplicon contamination and allows monitoring of amplification and analysis of results, making it more attractive to diagnostic testing laboratories. The improved sample preparation steps and the TaqMan real-time PCR assay provide a robust, streamlined, and rapid analytical procedure for surveillance, outbreak response, and regulatory testing of foods for detection of C. cayetanensis.

An IRT Investigation of the Validity of Non-Patient Analogue Research Using the Beck Depression Inventory

1995 ◽  
Vol 11 (1) ◽  
pp. 14-20 ◽  
Author(s):  
Sean M. Hammond
Keyword(s):  
Differential Item Functioning ◽  
Beck Depression Inventory ◽  
Rating Scale ◽  
Latent Trait ◽  
Scale Model ◽  
Item Functioning ◽  
Two Samples ◽  
Patient Groups ◽  
Analogue Research ◽  
Rating Scale Model

This paper presents an IRT analysis of the Beck Depression Inventory which was carried out to assess the assumption of an underlying latent trait common to non-clinical and patient samples. A one parameter rating scale model was fitted to data drawn from a patient and non-patient sample. Findings suggest that while the BDI fits the model reasonably well for the two samples separately there is sufficient differential item functioning to raise serious duobts of the viability of using it analogously with patient and non-patient groups.

Comparison of diagnostic accuracy of 18F-FDG PET, 123I-IMT- and 99mTc-MIBI SPECT

2006 ◽  
Vol 45 (01) ◽  
pp. 49-56 ◽  
Author(s):  
N. Özdemir-Sahin ◽  
P. Hipp ◽  
W. Mier ◽  
M. Eisenhut ◽  
J. Debus ◽  
...  
Keyword(s):  
Diagnostic Accuracy ◽  
Fdg Pet ◽  
Tumour Progression ◽  
Roc Curves ◽  
Low Grade ◽  
Who Grade ◽  
Pet Tracer ◽  
Patient Groups ◽  
99Mtc Mibi ◽  
Mibi Spect

Summary Aim was to evaluates the diagnostic accuracy of the SPECTtracers 3-123I-α-methyl-L-tyrosine (IMT) and 99mTc(I)- hexakis(2-methoxyisobutylisonitrile) (MIBI) as well as the PET-tracer 2-18F-2-deoxyglucose (FDG) for detecting tumour progression in irradiated low grade astrocytomas (LGA). Patients, methods: We examined 91 patients (56 males; 35 females; 44.7 ± 11.5 years), initially suffering from histologically proven LGAs (mean WHO grade II) and treated by stereotactic radiotherapy (59.0 ± 4.6 Gy). On average 21.9 ± 11.2 months after radiotherapy, patients presented new Gd-DTPA enhancing lesions on MRI, which did not allow a differentiation between progressive tumour (PT) and non-PT (nPT) at this point of time. PET scans (n=82) were acquired 45 min after injection of 208 ± 32 MBq FDG. SPECT scans started 10 min after injection of 269 ± 73 MBq IMT (n=68) and 15 min after injection of 706 ± 63 MBq MIBI (n=34). Lesions were classified as PT and nPT based on prospective follow-up (clinically, MRI) for 17.2 ± 9.9 months after PET/SPECT. Lesion-to-normal ratios (L/N) were calculated using contra lateraly mirrored reference regions for the SPECT examinations and reference regions in the contra lateral grey (GM) and white matter (WM) for FDG PET. Ratios were evaluated by Receiver Operating Characteristic (ROC) analysis. Results: In the patient groups nPT and PT, L/N ratios for FDG (GS) were 0.6 ± 0.3 vs. 1.2 ± 0.5 (p = 0.003), for FDG (WS) 1.2 ± 0.4 vs. 2.6 ± 0.4 (p <0.001), for IMT 1.1 ± 0.1 vs. 1.8 ± 0.4 (p <0.001) and for MIBI 1.6 ± 0.7 vs. 2.6 ± 2.2 (p = 0.554). Areas under the non-parametric ROC-curves were: 0.738 ± 0.059 for FDG (GS), 0.790 ± 0.057 for FDG (WS), 0.937 ± 0.037 for IMT and 0.564 ± 0.105 for MIBI. Conclusion: MIBI-SPECT examinations resulted in a low accuracy and especially in a poor sensitivity even at modest specificity values. A satisfying diagnostic accuracy was reached with FDG PET. Using WM as reference region for FDG PET, a slightly higher AUC as compared to GM was calculated. IMT yielded the best ROC characteristics and the highest diagnostic accuracy for differentiating between PT and nPT in irradiated LGA.

Anticardiolipin Antibodies Are Elevated in HIV-1 Infected Haemophiliacs but Do Not Predict for Disease Progression

1989 ◽  
Vol 61 (01) ◽  
pp. 081-085 ◽  
Author(s):  
Simon Panzer ◽  
Christoph Stain ◽  
Hubert Hartl ◽  
Robert Dudczak ◽  
Klaus Lechner
Keyword(s):  
Normal Values ◽  
Anticardiolipin Antibodies ◽  
Haemophilia A ◽  
Serum Samples ◽  
Factor Viii Concentrates ◽  
Patient Groups ◽  
Anti Hiv ◽  
Hiv 1 ◽  
Cd4 Lymphocytes

SummaryLevels of anticardiolipin antibodies (ACA) were measured in 55 patients with haemophilia A in serum samples obtained in 1983 and in 1987. Twenty-one patients were negative for anti HIV-1 antibodies in 1983 and remained negative in 1987; 34 patients had anti HIV-1 antibodies in 1983; 17 of these latter patients remained asymptomatic, whereas 17 patients developed ARC or AIDS during the 4 years follow-up. Thirteen anti HIV-1 negative patients had elevated ACA levels in 1983; subsequently, a significant decrease was observed in all these subjects (p <0.001). All anti HIV-1 positive patients had elevated ACA levels in 1983; normal values were found in 9 patients in 1987. Yet, these changes were not significant (p >0.05). ACA levels were significantly higher in HIV-1 infected patients than in those without anti HIV-1 antibodies (p <0.05). There was no difference of ACA levels between the two anti HIV-1 positive patient groups, be it in 1983 or be it in 1987 (p >0.05). There was no correlation of ACA levels with serum IgG concentrations, CD4+ lymphocytes, or the consumption of factor VIII concentrates.

Plasma Prekallikrein Activity in Patients with Total Hip Arthroplasty

1987 ◽  
Vol 58 (04) ◽  
pp. 1040-1042
Author(s):  
J J M L Hoffmann ◽  
J H J P M Kortmann
Keyword(s):  
Total Hip Arthroplasty ◽  
Hip Arthroplasty ◽  
Hip Prosthesis ◽  
Contact System ◽  
Factor Xii ◽  
Total Hip ◽  
Activity Factor ◽  
Patient Groups ◽  
Kallikrein Activity

SummaryThe behaviour of the contact system was studied in 40 patients with total hip arthroplasty, by measuring plasma prekallikrein, spontaneous kallikrein activity and factor XII. In the literature it had been shown that patients with complications from this operation had decreased prekallikrein and increased kallikrein activity (M. Nakahara. Acta orthop scand 1982; 53: 591-6). In the present study, comprising patients with and without pain and proven loosening of the hip prosthesis, these findings could only partially be confirmed. Patients with a loosened prosthesis had significantly lower prekallikrein (mean 0.78 ± 0.28 U/ml; p <0.01) than patients without problems, but no detectable kallikrein activity in plasma. Patients with pain but no loosening had normal prekallikrein (1.04 ±0 0.26 U/ml) and also no demonstrable kallikrein activity. Factor XII was normal in all patient groups. It is concluded that decreased prekallikrein is limited to patients with a loosened hip prosthesis, with or without pain.

Sign in / Sign up

Export Citation Format

Share Document