Further evaluation of an updated PCR assay for the detection of Schistosoma mansoni DNA in human stool samples

This study aimed to assess standard and harsher nucleic acid extraction schemes for diagnostic helminth real-time PCR approaches from stool samples. A standard procedure for nucleic acid extraction from stool and a procedure including bead-beating as well as proteinase K digestion were compared with group-, genus-, and species-specific real-time PCR assays targeting helminths and nonhelminth pathogens in human stool samples. From 25 different in-house and commercial helminth real-time PCR assays applied to 77 stool samples comprising 67 historic samples and 10 external quality assessment scheme samples positively tested for helminths, higher numbers of positive test results were observed after bead-beating-based nucleic acid extraction for 5/25 (20%) real-time PCR assays irrespective of specificity issues. Lower cycle threshold values were observed for one real-time PCR assay after the standard extraction scheme, and for four assays after the bead-beating-based scheme. Agreement between real-time PCR results after both nucleic acid extraction strategies according to Cohen’s kappa ranged from poor to almost perfect for the different assays. Varying agreement was observed in eight nonhelminth real-time PCR assays applied to 67 historic stool samples. The study indicates highly variable effects of harsh nucleic acid extraction approaches depending on the real-time PCR assay used.

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Development of a TaqMan based real-time PCR assay for detection of Clonorchis sinensis DNA in human stool samples and fishes

Parasitology International ◽

10.1016/j.parint.2011.06.010 ◽

2012 ◽

Vol 61 (1) ◽

pp. 183-186 ◽

Cited By ~ 23

Author(s):

Xian-Quan Cai ◽

Hai-Qiong Yu ◽

Jian-Shan Bai ◽

Jian-Dong Tang ◽

Xu-Chu Hu ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Clonorchis Sinensis ◽

Pcr Assay ◽

Stool Samples ◽

Human Stool

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Detection Cell-Free DNA (cfDNA) Using Nested-PCR as a Diagnosis Tool for Human Fascioliasis Infection

Iranian Journal of Public Health ◽

10.18502/ijph.v49i6.3367 ◽

2020 ◽

Author(s):

Mojgan ARYAEIPOUR ◽

Bahram KAZEMI ◽

Arezoo BOZORGOMID ◽

Mahdi MOHEBALI ◽

Hakim AZIZI ◽

...

Keyword(s):

Nested Pcr ◽

Deoxyribonucleic Acid ◽

Urine Samples ◽

Parasitic Diseases ◽

Conventional Pcr ◽

Pcr Assay ◽

Serum Samples ◽

Human Fascioliasis ◽

Stool Samples ◽

Human Stool

Background: We aimed to detect Fasciola specific deoxyribonucleic acid (DNA) by nested-PCR assay on human stool and urine samples and compare the results with the respective ELISA diagnostic assay. Methods: Overall, 206 clinically suspected cases of fascioliasis were enrolled in the study. Blood samples were collected from all the patients, and serum samples were isolated. ELISA assay, using Fasciola somatic antigen (SA), was carried out to detect anti Fasciola antibodies for the collected sera. DNA was randomly extracted from 25 stool and 10 urine samples of seropositive individuals and was evaluated by conventional PCR and nested PCR methods. The nested-PCR results were confirmed by sequencing the 430 bp region of ribosomal ITSI gene. Stool and urine samples from patients with different parasitic diseases and 25 stool samples from healthy individuals served as controls. Urine samples were collected from 10 healthy controls as well. Results: Fascioliasis was detected by ELISA in 24.8% of the individuals. Of these, 25 seropositive patients were randomly assigned to the study. Fasciola DNA was identified in the stool samples of 96% of seropositive patients by nested PCR but ova of Fasciola was detected by parasitology methods in only 20% of seropositive cases. Fasciola DNA was identified in 90% of the urine samples by nested PCR. No cross-reactions were observed with other parasites. Conclusion: Detection of cfDNA in stool and urine samples has high accuracy and thus can be used for the diagnosis of Fasciola infection in human.

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Development and Evaluation of a Real-Time PCR Assay for Detection and Quantification of Blastocystis Parasites in Human Stool Samples: Prospective Study of Patients with Hematological Malignancies

Journal of Clinical Microbiology ◽

10.1128/jcm.01392-10 ◽

2010 ◽

Vol 49 (3) ◽

pp. 975-983 ◽

Cited By ~ 91

Author(s):

P. Poirier ◽

I. Wawrzyniak ◽

A. Albert ◽

H. El Alaoui ◽

F. Delbac ◽

...

Keyword(s):

Prospective Study ◽

Real Time ◽

Real Time Pcr ◽

Hematological Malignancies ◽

Pcr Assay ◽

Stool Samples ◽

Human Stool ◽

Detection And Quantification

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Faculty Opinions recommendation of An in-depth analysis of a piece of shit: distribution of Schistosoma mansoni and hookworm eggs in human stool.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717972968.793469770 ◽

2013 ◽

Author(s):

Joachim Claudet

Keyword(s):

Schistosoma Mansoni ◽

Depth Analysis ◽

Human Stool

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Multicenter Evaluation of an ELISA for the Detection of Cryptosporidium spp. Antigen in Clinical Human Stool Samples

Microorganisms ◽

10.3390/microorganisms9020209 ◽

2021 ◽

Vol 9 (2) ◽

pp. 209

Author(s):

Romy Razakandrainibe ◽

Célia Mérat ◽

Nathalie Kapel ◽

Marc Sautour ◽

Karine Guyot ◽

...

Keyword(s):

Large Scale ◽

Cryptosporidium Oocyst ◽

Clinical Importance ◽

Epidemiological Studies ◽

Elisa Test ◽

Enzyme Linked Immunosorbent Assay ◽

Cryptosporidium Spp ◽

Stool Samples ◽

Conventional Microscopy ◽

Human Stool

Human cryptosporidiosis remains underdiagnosed, and rapid/accurate diagnosis is of clinical importance. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive, time-consuming, and requires skillful experience. Thus, we aimed to evaluate the usefulness of a coproantigen enzyme-linked immunosorbent assay (ELISA) test in detecting Cryptosporidium spp. from fecal specimens. For this aim, we evaluated the performances of a commercial ELISA (CoproELISA Cryptosporidium kit, Savyon Diagnostics, Israel) for the detection of Cryptosporidium spp. in random clinical stool samples through a multicenter study. The sensitivity and specificity for coproantigen ELISA were 98.86% and 94.32%, respectively. The coproantigen ELISA results indicate that the simple, rapid, reliable, and standardized immunoassay test is sensitive and specific for routine diagnosis, and may be useful for large-scale epidemiological studies of cryptosporidiosis.

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