Molecular characterization of Blastocystis isolates from birds by PCR with diagnostic primers and restriction fragment length polymorphism analysis of the small subunit ribosomal RNA gene

Abe, Niichiro; Wu, Zhiliang; Yoshikawa, Hideki

doi:10.1007/s00436-002-0782-5

Cited by 50 publications

(14 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similarly, in the NORMAT study, risk factors included contact with pigs or poultry within 7 days prior to stool submission. This is not surprising considering accumulating data from molecular studies of Blastocystis from potential reservoir hosts [3][4][5][34][35][36][37]. No particular subtype was associated with pig or poultry contact, and individuals reporting such contact were shown to host ST1, ST2, ST3 and/or ST4, which are the subtypes usually isolated from humans.…”

Section: Discussionmentioning

confidence: 97%

Blastocystis: unravelling potential risk factors and clinical significance of a common but neglected parasite

et al. 2009

View full text Add to dashboard Cite

Two independent studies were conducted to describe symptoms and potential risk factors associated with Blastocystis infection. Isolates were subtyped by molecular analysis. In the NORMAT study (126 individuals randomly sampled from the general population) 24 (19%) were positive for Blastocystis. Blastocystis was associated with irritable bowel syndrome (P=0.04), contact with pigs (P<0.01) and poultry (P=0.03). In the Follow-up (FU) study (follow-up of 92 Blastocystis-positive patients), reports on bloating were associated with subtype (ST) 2 (P<0.01), and blood in stool to mixed subtype infection (P=0.06). ST1 was more common in FU individuals (32%) than in NORMAT individuals (8%), whereas single subtype infections due to ST3 or ST4 were seen in 63% of the NORMAT cases and 28% of the FU cases. Only FU individuals hosted ST7, and ST6/7 infections due to ST7 or ST9 were characterized by multiple intestinal symptoms. The data indicate subtype-dependent differences in the clinical significance of Blastocystis.

show abstract

Section: Discussionmentioning

confidence: 97%

Blastocystis: unravelling potential risk factors and clinical significance of a common but neglected parasite

et al. 2009

View full text Add to dashboard Cite

show abstract

“…PCR was carried out with specific sequence-tagged site (STS) primers (Table 1). Seven different STS primers were used for genotyping as described previously [6,7,12-18]. The STS primers used were SB83 (351 bp), SB340 (704 bp), SB227 (526 bp), SB337 (487 bp), SB336 (317 bp), SB332 (338 bp) and SB 155 (650 bp) for subtype 1, 2, 3, 4, 5, 6 and 7 respectively based on a recent classification terminology [19].…”

Section: Methodsmentioning

confidence: 99%

Advantage of using colonic washouts for Blastocystis detection in colorectal cancer patients

et al. 2014

View full text Add to dashboard Cite

BackgroundThere have been previous studies associating microorganisms to cancer and with our recent findings of Blastocytsis antigen having a higher in vitro proliferation of cancer cells strengthens the suspicion. Collecting faecal samples alone to associate this parasite with cancer may not be accurate due to the phenomenon of irregular shedding and the possible treatment administrated to the cancer patients. Hence, this become the basis to search for an alternate method of sample collection. Colonic washout is an almost complete washed up material from colon and rectum which includes various microorganisms such as Blastocystis and other lodged material within the villi. The detection of parasite in colonic washouts will give a better reflection on the association between Blastocystis and CRC.MethodsBlastocytsis detection was made by in vitro culture method using Jones’ medium, formal ether concentration technique and conventional polymerase chain reaction (PCR) on faecal samples and colonic washouts of 204 CRC patients from colonoscopy procedure. Faecal samples and colonic washouts from 221 normal individuals served as control.ResultsWe observed an increased detection of Blastocystis using colonic washouts (n = 53, 12.47%) than faecal samples (n = 26, 6.12%). Eleven faecal samples showed positive results for Blastocystis which were also found in colonic washouts using the PCR technique. This study for the first time showed a significant Blastocystis infection among CRC patients (n = 43, 21.08%) compared to the asymptomatic normal individuals (n = 22, 9.95%). Blastocystis subtype 3 infection was found to be significantly more prevalent (n = 26, 12.75%) compared to other subtypes namely subtype 1: n = 9 (4.41%), subtype 2: n = 1 (0.49%), subtype 5: n = 1 (0.49%) and mixed subtype: n = 6 (2.94%) among the CRC patients.ConclusionThe study showed that colonic washouts provide a better alternative for Blastocystis detection in CRC patients compared to faecal samples as this prevents treatment regime and the phenomenon of irregular shedding from influencing the detection results obtained from faecal samples.

show abstract

“…showing the different RFLP patterns of various isolates of Blastocystis spp. [42,43,44,45,46]. However, only some studies report PCR-RFLP analysis with different primers and restriction enzymes to identify subtypes of Blastocystis spp.…”

Section: Introductionmentioning

confidence: 99%

A Simple Genotyping Method for Rapid Differentiation of Blastocystis Subtypes and Subtype Distribution of Blastocystis spp. in Thailand

et al. 2019

View full text Add to dashboard Cite

Blastocystis spp. is one of the most common protozoa of humans and animals worldwide. The genetic diversity of Blastocystis spp. might be associated with a wide range of symptoms. However, the prevalence of each subtype is different in each country. Until now, there is no standard method for subtyping of Blastocystis spp. We developed a sequential restriction fragment length polymorphism (RFLP) analysis for the rapid differentiation of human Blastocystis subtypes. A large-scale study was also conducted to determine the subtype distribution of Blastocystis spp. in Thailand. Stool samples were collected from 1025 school-age students in four regions of Thailand. Blastocystis infections were identified by direct smear, formalin ethyl-acetate concentration technique (FECT), Boeck and Drbohlav’s Locke-Egg-Serum (LES) medium culture, and polymerase chain reaction (PCR) of small-subunit ribosomal DNA (SSU rDNA). Subtypes of Blastocystis spp. were determined by RFLP. Phylogenetic tree of partial SSU rDNA sequences of Blastocystis spp. was constructed using the Maximum Likelihood (ML) method. Out of 1025 students, 416 (40.6%) were positive for Blastocystis spp. Using two steps of RFLP reactions, we could determine subtype one–three among these students. Subtype 3 was the most common subtype (58.72%) in Thai students, followed by subtype 1 (31.2%), and subtype 2 (10.1%). Blastocystis subtype 3 was the most prevalent in all regions of Thailand. The subtype distribution of Blastocystis spp. in Thailand was different from other countries.

show abstract

Molecular characterization of Blastocystis isolates from birds by PCR with diagnostic primers and restriction fragment length polymorphism analysis of the small subunit ribosomal RNA gene

Cited by 50 publications

References 23 publications

Blastocystis: unravelling potential risk factors and clinical significance of a common but neglected parasite

Blastocystis: unravelling potential risk factors and clinical significance of a common but neglected parasite

Advantage of using colonic washouts for Blastocystis detection in colorectal cancer patients

A Simple Genotyping Method for Rapid Differentiation of Blastocystis Subtypes and Subtype Distribution of Blastocystis spp. in Thailand

Contact Info

Product

Resources

About