scholarly journals Commercial Simplex and Multiplex PCR Assays for the Detection of Intestinal Parasites Giardia intestinalis, Entamoeba spp., and Cryptosporidium spp.: Comparative Evaluation of Seven Commercial PCR Kits with Routine In-House Simplex PCR Assays

2021 ◽  
Vol 9 (11) ◽  
pp. 2325
Author(s):  
Louise Basmaciyan ◽  
Alexandre François ◽  
Anne Vincent ◽  
Stéphane Valot ◽  
Alain Bonnin ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Intestinal Parasites ◽  
Dna Amplification ◽  
Giardia Intestinalis ◽  
Cryptosporidium Spp ◽  
Stool Samples ◽  
Pcr Assays ◽  
Commercial Kits ◽  
Sensitivity Specificity ◽  
Interesting Alternative

Nowadays, many commercial kits allowing the detection of digestive parasites by DNA amplification methods have been developed, including simplex PCR assays (SimpPCRa) allowing the identification of a single parasite, and multiplex PCR assays (MultPCRa) allowing the identification of several parasites at once. Thus, aimed at improving the diagnosis of intestinal protozoal infections, it is essential to evaluate the performances of these new tools. A total of 174 DNA samples collected between 2007 and 2017 were retrospectively included in this study. Performances of four commercial SimpPCRa (i.e., CerTest-VIASURETM) and three MultPCRa (i.e., CerTest-VIASURETM, FAST-TRACK-Diagnostics-FTD-Stool-ParasiteTM and DIAGENODE-Gastroenteritis/Parasite-panel-ITM) were evaluated for the detection of Cryptosporidium spp., Entamoeba spp., and Giardia intestinalis in stool samples compared to our routinely used in-house SimpPCRa. Globally, the SimpPCRa showed better sensitivity/specificity for the detection of G. intestinalis, E. histolytica, E. dispar, and Cryptosporidium spp. (i.e., 96.9/93.6%; 100/100%; 95.5/100%; and 100/99.3%, respectively), compared to the three commercial MultPCRa tested. All in all, we showed that MultPCRa offer an interesting alternative for the detection of protozoans in stool samples depending on the clinical context.

scholarly journals Selecting PCR for the Diagnosis of Intestinal Parasitosis: Choice of Targets, Evaluation of In-House Assays, and Comparison with Commercial Kits

2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
G. N. Hartmeyer ◽  
S. V. Hoegh ◽  
M. N. Skov ◽  
R. B. Dessau ◽  
M. Kemp
Keyword(s):  
Intestinal Parasites ◽  
Parasitic Infection ◽  
Rt Pcr ◽  
Specific Pcr ◽  
Intestinal Parasitosis ◽  
Stool Samples ◽  
Pcr Assays ◽  
Specific Pcr Assay ◽  
Commercial Kits ◽  
Almost All

Microscopy of stool samples is a labour-intensive and inaccurate technique for detection of intestinal parasites causing diarrhoea and replacement by PCR is attractive. Almost all cases of diarrhoea induced by parasites over a nine-year period in our laboratory were due toGiardia lamblia,Cryptosporidiumspecies, orEntamoeba histolyticadetected by microscopy. We evaluated and selected in-house singleplex real-time PCR (RT-PCR) assays for these pathogens in 99 stool samples from patients suspected of having intestinal parasitosis tested by microscopy. The strategy included a genus-specific PCR assay forC. parvumandC. hominis, with subsequent identification by a PCR that distinguishes between the two species.G. lambliawas detected in five andC. parvumin one out of 68 microscopy-negative samples. The performance of the in-house RT-PCR assays was compared to three commercially available multiplex test (MT-PCR) kit systems in 81 stool samples, collected in 28 microscopy-positive and 27 microscopy-negative samples from individuals suspected of intestinal parasitosis and in 26 samples from individuals without suspicion of parasitic infection. The in-house assays detected parasites in more samples from patients suspected of having parasitosis than did any of the kits. We conclude that commercial kits are targeting relevant parasites, but their performance may vary.

scholarly journals Detection of Cryptosporidium spp and other intestinal parasites in children with acute diarrhea and severe dehydration in Rio de Janeiro

2007 ◽  
Vol 40 (3) ◽  
pp. 346-348 ◽  
Author(s):  
Filipe Anibal Carvalho-Costa ◽  
Alessandra Queiroga Gonçalves ◽  
Sandra Laranjeira Lassance ◽  
Carla Pontes de Albuquerque ◽  
José Paulo Gagliardi Leite ◽  
...  
Keyword(s):  
Giardia Lamblia ◽  
Intestinal Parasites ◽  
Blue Staining ◽  
Pediatric Hospital ◽  
Severe Dehydration ◽  
Enterobius Vermicularis ◽  
Blastocystis Hominis ◽  
Entamoeba Dispar ◽  
Cryptosporidium Spp ◽  
Stool Samples

The objective of the present study was to estimate the frequency of infection by Cryptosporidium spp and other intestinal parasites in dehydrated children with gastroenteritis who were admitted to a pediatric hospital. Stool examinations from 218 children were performed. Cryptosporidium spp was identified in eighteen out of 193 stool samples (9.3%) subjected to safranin-methylene blue staining. Giardia lamblia was detected in ten out of 213 (4.7%) samples examined via the direct or Ritchie methods. Other parasites identified were Ascaris lumbricoides (4.2%), Blastocystis hominis (1.4%), Entamoeba coli (0.9%), Entamoeba histolytica/Entamoeba dispar (0.5%), Endolimax nana (0.5%), Trichuris trichiura (0.5%) and Enterobius vermicularis (0.5%).

scholarly journals Diagnosis of Blastocystis sp., Cryptosporidium sp., and Giardia intestinalis by Multiplex PCR: An Optimization Study

2021 ◽  
Author(s):  
Ali Ahmet Kilimcioğlu ◽  
Nogay Girginkardeşler ◽  
Tuba Oyur ◽  
Selin Bölük Sabuncu ◽  
Didem Düzyol Azak ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Dna Isolation ◽  
Giardia Intestinalis ◽  
Molecular Method ◽  
Chain Reaction ◽  
Optimization Study ◽  
Optimized Protocol ◽  
Blastocystis Sp ◽  
Stool Samples ◽  
Polymerase Chain

Objective: It was aimed to develop a new Multiplex Polymerase Chain Reaction (PCR) protocol with isolates obtained from local patients for the diagnosis of Blastocystis sp., Cryptosporidium sp. and Giardia intestinalis, which can cause severe gastrointestinal system complaints especially in immunocompromised patients and children. Method: DNA isolation was performed with a commercial kit from three stool samples of different patients whose microscopic examination showed dense amounts of Blastocystis sp., Cryptosporidium sp. and Giardia intestinalis. First, a special PCR protocol has been developed for each protozoon. Then, the multiplex PCR protocol, in which these three protozoa can be diagnosed together, was optimized. Results: In the multiplex PCR protocol performed after DNA isolation, bands of 95 bp., 227 bp. and 258 bp. were obtained for Cryptosporidium sp., Blastocystis sp. and G. intestinalis, respectively. Conclusion: Blastocystis sp., Cryptosporidium sp. and Giardia intestinalis were diagnosed by multiplex PCR with the original protocol developed. Due to the difficulties in using different methods in parasitological examination, by adding other protozoa important for public health to this optimized protocol, it will be possible to detect a large number of parasites with a single molecular method.

scholarly journals Evaluation of the AllplexTM Gastrointestinal Panel—Parasite Assay for Protozoa Detection in Stool Samples: A Retrospective and Prospective Study

2020 ◽  
Vol 8 (4) ◽  
pp. 569 ◽  
Author(s):  
Brice Autier ◽  
Jean-Pierre Gangneux ◽  
Florence Robert-Gangneux
Keyword(s):  
Prospective Study ◽  
Multiplex Pcr ◽  
Clinical Samples ◽  
Molecular Assay ◽  
Pcr Assay ◽  
Cryptosporidium Spp ◽  
Multiplex Pcr Assay ◽  
Parasite Detection ◽  
Stool Samples ◽  
Higher Sensitivity

This study aims at evaluating the performances of the multiplex PCR AllplexTM Gastrointestinal Panel-Parasite Assay (GIPPA), which detects G. duodenalis, Cryptosporidium spp., E. histolytica, D. fragilis, B. hominis, and C. cayetanensis, by comparison to microscopy. A retrospective evaluation was conducted on a series of positive clinical samples (n = 99) stored at −80 °C or at +4 °C. A five-month prospective study was then conducted on all samples sent to our lab for parasite detection (n = 586). In the retrospective cohort, sensitivity was 81% for both G. duodenalis (26/32) and D. fragilis (21/26) and 100% for Cryptosporidium spp. (26/26, including 6 different species), B. hominis (26/26), and C. cayetanensis (4/4). During the prospective study, 95 samples were positive by microscopy and 207 by multiplex PCR assay. The molecular assay showed a significantly higher sensitivity of PCR, especially for G. duodenalis (100% vs. 60.7%, p < 0.01), D. fragilis (97.2% vs. 14.1%, p < 0.001), and B. hominis (99.4% vs. 44.2%, p < 0.001) but also for E. histolytica (100% vs. 50.0%). The sensitivity of the AllplexTM GIPPA on the first stool sample was equivalent to the sensitivity of microscopy on multiple stool samples but inferior to multiplex PCR on multiple stool samples. Taken together, the AllplexTM GIPPA is suitable for the routine detection of protozoa in fecal samples.

scholarly journals Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths

Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 152 ◽  
Author(s):  
Vivornpun Sanprasert ◽  
Ruthairat Kerdkaew ◽  
Siriporn Srirungruang ◽  
Sarit Charuchaibovorn ◽  
Kobpat Phadungsaksawasdi ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Real Time ◽  
Real Time Pcr ◽  
Intestinal Parasites ◽  
Simultaneous Detection ◽  
Diagnostic Tools ◽  
Multiple Infections ◽  
Parasitic Infections ◽  
Soil Transmitted Helminths ◽  
Stool Samples

Soil-transmitted helminths (STHs) are the most common intestinal parasites infecting humans worldwide. STH infections are a major cause of morbidity and disability. Accurate diagnostic tools are pivotal for assessing the exact prevalence of parasitic infections. Microscopic examination and culture techniques have been used to observe the presence of eggs or larvae of parasites in stool samples, but they are time-consuming and have low sensitivity. Therefore, accurate, simple, and inexpensive diagnostic techniques are still required for simultaneous detection of STH infections. Although molecular-based techniques, such as real-time PCR and multiplex real-time PCR, have been developed, they are not suitable for routine diagnosis due to the requirement for expensive reagents and instruments. In this study, we established a conventional multiplex PCR for simultaneous rapid detection of Ascaris lumbricoides, Necator americanus, and Strongyloides stercoralis in stool samples. Our results show that the multiplex PCR could detect the DNA of STHs at a very low target gene concentrations (lower than 1 pg) with no cross-amplification. Multiplex PCR had five times higher sensitivity than the formalin–ethyl acetate concentration technique (FECT) in the detection of multiple infections, and two times higher for detection of S. stercoralis. However, multiplex PCR was comparable to FECT in the detection of A. lumbricoides and N. americanus. In conclusion, this method could be used as an alternative method for the detection of STHs, especially for S. stercoralis.

scholarly journals Prevalence of Giardia intestinalis Infection in Schistosomiasis-Endemic Areas in South-Central Mali

2019 ◽  
Vol 4 (2) ◽  
pp. 86 ◽  
Author(s):  
Hassan K.M. Fofana ◽  
Maren Schwarzkopf ◽  
Mama N. Doumbia ◽  
Rénion Saye ◽  
Anna Nimmesgern ◽  
...  
Keyword(s):  
Intestinal Parasites ◽  
Epidemiological Data ◽  
Giardia Intestinalis ◽  
Diagnostic Tools ◽  
Intestinal Protozoa ◽  
Control Programs ◽  
South Central ◽  
Concurrent Infections ◽  
Intestinal Pathogens ◽  
Stool Samples

Intestinal parasite infections are frequent causes of diarrhea and malnutrition among children in the tropics. Transmission of helminths and intestinal protozoa is intimately connected with conditions of poverty, including inadequate sanitation and hygiene. Concurrent infections with several intestinal pathogens may lead to excess morbidity. Yet, there is a paucity of epidemiological data from Mali. In this study, stool samples from 56 individuals, aged 2–63 years, from Bamako and Niono, south-central Mali were examined for intestinal parasites using stool microscopy. Additionally, stool samples were subjected to a rapid diagnostic test (RDT) and polymerase chain reaction (PCR) for the detection of Cryptosporidium spp. and Giardia intestinalis. The predominant pathogens were Schistosoma mansoni and G. intestinalis with prevalences of 41% and 38%, respectively. Hymenolepis nana was detected in 4% of the participants, while no eggs of soil-transmitted helminths were found. Concurrent infections with G. intestinalis and S. mansoni were diagnosed in 16% of the participants. For the detection of G. intestinalis, PCR was more sensitive (100%) than RDT (62%) and microscopy (48%). As helminth-protozoa coinfections might have important implications for morbidity control programs, future studies should employ diagnostic tools beyond stool microscopy to accurately assess the co-endemicity of giardiasis and schistosomiasis.

scholarly journals Prevalence of intestinal parasites in children from minority group with low hygienic standards in Slovakia

2012 ◽  
Vol 49 (2) ◽  
pp. 63-66 ◽  
Author(s):  
P. Rudohradská ◽  
M. Halánová ◽  
P. Ravaszová ◽  
M. Goldová ◽  
A. Valenčáková ◽  
...  
Keyword(s):  
Socioeconomic Status ◽  
Human Population ◽  
Intestinal Parasites ◽  
Minority Group ◽  
Developed Countries ◽  
Giardia Intestinalis ◽  
Protozoan Parasites ◽  
Cryptosporidium Spp ◽  
Faecal Samples ◽  
Hygienic Standard

AbstractThe number of parasites followed the rapid growing of human population worldwide, not only in developing but also in developed countries. Many of them are diagnosed in children and adolescents. The occurrence of selected intestinal endoparasites in children coming from areas with low hygienic and socioeconomic status was studied. Out of 81 faecal samples examined, 46 (56.8 %) were positive for presence of intestinal parasites. From helminths, Ascaris lumbricoides was found to be the leading parasite (24.7 %), followed by Trichuris trichiura (17.3 %). Tapeworm Taenia spp. eggs were detected in 4.9 % of examined children. From protozoan parasites Cryptosporidium spp. was observed in 36 children (44.4 %) and Giardia intestinalis in 20 children (24.7 %). The occurrence of these epidemiologically low risky parasites in Roma children population suggests low hygienic standard in the Roma settlements.

scholarly journals The effect of water source and soil supplementation on parasite contamination in organic vegetable gardens

2018 ◽  
Author(s):  
Fernanda Pinto Ferreira ◽  
Eloiza Teles Caldart ◽  
Roberta Lemos Freire ◽  
Regina Mitsuka-Breganó ◽  
Felipe Machado de Freitas ◽  
...  
Keyword(s):  
Management Practices ◽  
Dna Amplification ◽  
Water Source ◽  
Giardia Intestinalis ◽  
Chain Reaction ◽  
Factors Associated ◽  
Cryptosporidium Spp ◽  
Etiologic Agents ◽  
Vegetable Gardens ◽  
Polymerase Chain

Abstract The objective of this study was to determine factors associated with vegetable contamination with zoonotic protozoan. Samples of water, soil and vegetables were collected from July/2014 to May/2016, totaling 83 samples, 21 properties of Londrina region, Paraná, Brazil. DNA amplification of Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis in the samples was conducted using polymerase chain reaction (PCR). The PCR results were positive for T. gondii in 12.9% (8/62), Cryptosporidium spp. in 11.3% (7/62) and G. intestinalis in 25.8% (16/62) of the samples. DNA sequencing identified C. parvum in five samples and G. intestinalis Assemblage E in three. The statistical associations demonstrated greater probability of positive samples for T. gondii and for at least one of the three protozoa when the source of irrigation water was the river; a greater chance of positive samples for Cryptosporidium spp. when deer were present on the property; and a smaller chance of positive samples for at least one of the three etiologic agents when soil was supplemented with limestone. The results expose some critical contamination points, providing support for training farmers on good management practices during the production process.

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