Genetic characterization of Toxoplasma gondii in Iranian HIV positive patients using multilocus nested-PCR-RFLP method

Parasitology ◽  
2019 ◽  
Vol 147 (3) ◽  
pp. 322-328
Author(s):  
Seyed Abdollah Hosseini ◽  
Mehdi Sharif ◽  
Shahabeddin Sarvi ◽  
Saeid Abediankenari ◽  
Mohammad Bagher Hashemi-Soteh ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hiv Positive ◽  
Type I ◽  
Serum Samples ◽  
Nested Polymerase Chain Reaction ◽  
Northern Iran ◽  
Type Iii ◽  
Genotype 2 ◽  
Pcr Rflp

AbstractThe aim of the present study was to investigate the prevalence and genotyping of Toxoplasma gondii in Iranian human immunodeficiency virus (HIV)-positive patients using multilocus-nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP). A total of 102 serum samples obtained from infected patients were collected from the laboratory centres in northern Iran. Anti-T. gondii antibodies and deoxyribonucleic acid (DNA) detection were accomplished by an enzyme-linked immunosorbent assay and PCR. The Mn-PCR-RFLP method was used for the genotyping of T. gondii. Overall, 68.6% (70/102) and 11.7% (12/102) of the individuals were tested positive for anti-T. gondii immunoglobulin G and T. gondii DNA, respectively. Complete genotyping was performed on 10/12 (83.3%) PCR-positive samples. Accordingly, the samples were classified as genotype #1 (type II clonal; n = 3, 30%), genotype #2 (type III clonal; n = 2, 20%), genotype #10 (type I clonal; n = 2, 20%), genotype #27 (type I variant; n = 1, 10%), genotype #35 (type I variant; n = 1, 10%) and genotype #48 (type III variant; n = 1, 10%). The results were indicative of the high frequency of the type I and type I variant of T. gondii strains in HIV-positive patients in northern Iran. Given the high prevalence of T. gondii and frequency of pathogenic types (pathogen in laboratory mice) in the patients, special measures should be taken to prevent the possible increased incidence of encephalitis by T. gondii.

Toxoplasmosis among cancer patients undergoing chemotherapy: a population study based on the serological, molecular and epidemiological aspects

2020 ◽  
Author(s):  
Seyed Abdollah Hosseini ◽  
Mehdi Sharif ◽  
Shahabeddin Sarvi ◽  
Ghasem Janbabai ◽  
Shahrbanoo Keihanian ◽  
...  
Keyword(s):  
Cancer Patients ◽  
Fragment Length Polymorphism ◽  
Type I ◽  
Type Ii ◽  
Genotype 1 ◽  
Serum Samples ◽  
Northern Iran ◽  
Patients With Cancer ◽  
Genotype 2 ◽  
High Prevalence

Abstract Background Toxoplasmosis is highly prevalent in northern Iran and immunocompromised individuals are more susceptible to this infection. The present study aimed to determine the seroprevalence, parasitism and genetic diversity of Toxoplasma gondii among patients with cancer undergoing chemotherapy in northern Iran. Methods A total of 350 serum samples obtained from cancer patients were collected from laboratory centers in northern Iran. Immunodiagnosis and DNA detection were accomplished by ELISA and PCR. Thereafter, multiplex-nested PCR-restriction fragment length polymorphism was used for the genotyping of T. gondii. Results In general, out of 350 patients, 264 (75.4%) and 9 (2.57%) cases were positive for anti-T. gondii IgG and IgM, respectively. Moreover, 19 (5.43%) samples contained T. gondii DNA. From 19 positive samples, 10 high-quality samples with sharp and non-smear bands were selected to determine the genotypes of T. gondii. Accordingly, the samples were classified as genotype #1 (type II clonal; n=4, 40%), genotype #2 (type III clonal; n=3, 30%), genotype #10 (type I clonal; n=2, 20%) and genotype #27 (type I variant; n=1, 10%). Conclusions As evidenced by the results, due to the high prevalence of T. gondii, cancer patients in northern Iran are at serious risk of severe toxoplasmosis and its complications. Therefore, oncologists need to regard this critical health problem as a matter requiring urgent attention.

scholarly journals Use of GRA6-Derived Synthetic Polymorphic Peptides in an Immunoenzymatic Assay To Serotype Toxoplasma gondii in Human Serum Samples Collected from Three Continents

2008 ◽  
Vol 15 (9) ◽  
pp. 1380-1386 ◽  
Author(s):  
Susana Sousa ◽  
Daniel Ajzenberg ◽  
Manuel Vilanova ◽  
José Costa ◽  
Marie-Laure Dardé
Keyword(s):  
Toxoplasma Gondii ◽  
Latin American ◽  
Enzyme Linked Immunosorbent Assay ◽  
Clear Correlation ◽  
Type I ◽  
Serum Samples ◽  
Typing Method ◽  
Microsatellite Genotype ◽  
Human Infections ◽  
Immunoenzymatic Assay

ABSTRACT Serotyping is a simple typing method that consists of an immunoenzymatic assay (enzyme-linked immunosorbent assay [ELISA]) using synthetic polymorphic peptides derived from Toxoplasma gondii antigens. We developed a new ELISA based on GRA6 C-terminal polymorphic peptides. Serum samples from 41 human infections due to 23 archetypal (type I, II, or III) and 18 nonarchetypal strains were selected in order to validate this approach. For 20 out of the 23 archetypal infections, there was a clear correlation between microsatellite genotype and GRA6 serotyping. All infections due to nonarchetypal strains were misclassified as archetypal strain infections. The GRA6 C-terminal peptides from these strains were analyzed to explain this misclassification. A second group of 455 patients with acute and chronic toxoplasmosis due to unknown genotypes from different European, African, and Latin American countries were included in this study, and the strain type predicted by this method. The results suggest that serotyping is a promising method for typing strains, although limitations exist for African and South American strains as a consequence of higher peptide polymorphism. Other peptides from different markers must be studied in order to discriminate archetypal from nonarchetypal strains.

scholarly journals Detection and genetic characterisation of Toxoplasma gondii circulating in free-range chickens, pigs and seropositive pregnant women in Benue state, Nigeria

2021 ◽  
Vol 15 (6) ◽  
pp. e0009458
Author(s):  
Ifeoma N. Nzelu ◽  
Jacob K. P. Kwaga ◽  
Junaidu Kabir ◽  
Idris A. Lawal ◽  
Christy Beazley ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Toxoplasma Gondii ◽  
Pregnant Women ◽  
Quantitative Pcr ◽  
Human Consumption ◽  
Type I ◽  
Nested Polymerase Chain Reaction ◽  
Free Range ◽  
Animal Populations ◽  
Pcr Rflp

Toxoplasma gondii parasites present strong but geographically varied signatures of population structure. Populations sampled from Europe and North America have commonly been defined by over-representation of a small number of clonal types, in contrast to greater diversity in South America. The occurrence and extent of genetic diversity in African T. gondii populations remains understudied, undermining assessments of risk and transmission. The present study was designed to establish the occurrence, genotype and phylogeny of T. gondii in meat samples collected from livestock produced for human consumption (free-range chickens, n = 173; pigs, n = 211), comparing with T. gondii detected in blood samples collected from seropositive pregnant women (n = 91) in Benue state, Nigeria. The presence of T. gondii DNA was determined using a published nested polymerase chain reaction, targeting the 529 bp multicopy gene element. Samples with the highest parasite load (assessed using quantitative PCR) were selected for PCR-restriction fragment length polymorphism (PCR-RFLP) targeting the surface antigen 3 (SAG3), SAG2 (5’ and 3’), beta-tubulin (BTUB) and dense granule protein 6 (GRA6) loci, and the apicoplast genome (Apico). Toxoplasma gondii DNA was detected in all three of the populations sampled, presenting 30.6, 31.3 and 25.3% occurrence in free-range chickens, pigs and seropositive pregnant women, respectively. Quantitative-PCR indicated low parasite occurrence in most positive samples, limiting some further molecular analyses. PCR-RFLP results suggested that T. gondii circulating in the sampled populations presented with a type II genetic background, although all included a hybrid type I/II or II/III haplotype. Concatenation of aligned RFLP amplicon sequences revealed limited diversity with nine haplotypes and little indication of host species-specific or spatially distributed sub-populations. Samples collected from humans shared haplotypes with free-range chickens and/or pigs. Africa remains under-explored for T. gondii genetic diversity and this study provides the first detailed definition of haplotypes circulating in human and animal populations in Nigeria.

Genetic characterization of Toxoplasma gondii isolates from chickens in India by GRA6 gene sequence analysis

2014 ◽  
Vol 59 (4) ◽  
Author(s):  
Shantaveer Biradar ◽  
Buddhi Saravanan ◽  
Anup Tewari ◽  
Chirukandoth Sreekumar ◽  
Muthu Sankar ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Toxoplasma Gondii ◽  
Amino Acid Sequences ◽  
Nucleotide Sequencing ◽  
Type I ◽  
Type Iii ◽  
Pcr Rflp ◽  
Indian Isolates ◽  
Gra6 Gene

AbstractPCR-RFLP and nucleotide sequencing based genotyping of Toxoplasma gondii Indian isolates (Izatnagar and Chennai isolates and Chennai clone) vis-a vis RH-IVRI strain was conducted by targeting GRA6 as genetic marker. The 791 bp GRA6 product was PCR amplified from the genomic DNA of different T. gondii Indian isolates, including the RH-IVRI strain. Tru1I restriction endonuclease based PCR-RFLP of GRA6 sequence produced polymorphic digestion pattern that discriminated the virulent RH-IVRI strain (as type I) from the moderately virulent local isolates as type III. The PCR amplicon of T. gondii GRA6 from RH-IVRI strain as well as from the local isolates were cloned in cloning vector and custom sequenced. The nucleotide and deduced amino acid sequences of T. gondii isolates were aligned with that of the type I, II and III strains (RH, BEVERLEY, ME49, C56, TONT and NED) available in public domain and analyzed in silico using MEGA version 4.0 software. Nucleotide sequencing and phylogenetic analysis of GRA6 marker from the Indian isolates revealed a close genetic relationship with type III strains of T. gondii. Further, detection of a single nucleotide polymorphism (SNP) at positions 162 and 171 of the GRA6 marker, established the lineage of Indian isolates as type III. This is the first report on characterization of T. gondii lineage as type III in selective chicken population of India based on PCR-RFLP and sequence analysis of GRA6 gene.

scholarly journals Toxoplasma gondii genotypes circulating in domestic pigs in Serbia

2019 ◽  
Vol 67 (2) ◽  
pp. 204-211 ◽  
Author(s):  
Ljiljana Kuruca ◽  
Aleksandra Uzelac ◽  
Ivana Klun ◽  
Vesna Lalošević ◽  
Olgica Djurković-Djaković
Keyword(s):  
Toxoplasma Gondii ◽  
Significant Risk ◽  
Significant Risk Factor ◽  
Human Infection ◽  
Nested Polymerase Chain Reaction ◽  
Type Iii ◽  
Pcr Rflp ◽  
Slaughter Pigs ◽  
Commercial Farms ◽  
Dna Genotyping

Consumption of undercooked or raw pork is considered a significant risk factor for human infection with Toxoplasma gondii. In this study, we investigated the genetic structure of 18 T. gondii strains obtained from slaughter pigs from Northern Serbia (mainly Vojvodina). The examined samples originated from eight pigs from large commercial farms, six backyard pigs and four free-range Mangalica pigs, all found to be positive for either viable T. gondii or T. gondii DNA. Genotyping was attempted from both pig tissues and mouse brains from the bio-assays using a multiplex multilocus nested polymerase chain reaction–restriction fragment length polymorphism (Mn-PCR-RFLP) method with seven markers (GRA6, alt. SAG2, PK-1, BTUB, C22-8, CS3 and Apico). Identification was achieved for nine T. gondii isolates. Seven isolates were classified as type II and two as type III. These results are consistent with previous studies on animal isolates from Serbia as well as with previous reports that type III is more frequently found in samples from Southern Europe than in those from other parts of the continent.

scholarly journals Diagnosis and isolation of Toxoplasma gondii in horses from Brazilian slaughterhouses

2013 ◽  
Vol 22 (1) ◽  
pp. 58-63 ◽  
Author(s):  
Fernanda Evers ◽  
João Luis Garcia ◽  
Italmar Teodorico Navarro ◽  
Dauton Luiz Zulpo ◽  
Beatriz de Souza Lima Nino ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Neospora Caninum ◽  
Fluorescent Antibody ◽  
Rflp Analysis ◽  
Antibody Test ◽  
Mouse Bioassay ◽  
Type I ◽  
Serum Samples ◽  
Sarcocystis Neurona ◽  
Pcr Rflp

This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64), and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6%) by IFAT and in 14 (3.5%) by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ 1:16), T. gondii being isolated in the remaining two. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of 18S rDNA to differentiate among T. gondii, Neospora caninum, and Sarcocystis neurona, we found that two of the 14 brains were positive for T. gondii only. For genotyping of the two isolates and the PCR-positive brain, we performed PCR-RFLP based on 13 markers, and SAG2 all samples were Toxoplasma gondii type I. Collectively, IFAT of horse sera and mouse bioassay identified positivity in 60 (15%) of the samples. Our results show that some horses sent to slaughter in Brazil have been exposed toT. gondii.

scholarly journals Seroprevalence of Toxoplasma gondii and associated risk factors in domestic pigs raised from Cuba

2021 ◽  
Author(s):  
Julio César Castillo-Cuenca ◽  
Álvaro Martínez-Moreno ◽  
José Manuel Diaz-Cao ◽  
Angel Entrena-García ◽  
Jorge Fraga ◽  
...  
Keyword(s):  
Risk Factors ◽  
Toxoplasma Gondii ◽  
Meat Products ◽  
Control Measures ◽  
Health Concern ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Cross Sectional ◽  
Weaning Pigs ◽  
Associated Risk Factors

AbstractA cross-sectional study was carried out to determine the seroprevalence of Toxoplasma gondii and associated risk factors in pigs in the largest pork-producing region in Cuba. Serum samples from 420 pigs, including 210 sows and 210 post-weaning pigs, were tested for antibodies against T. gondii using a commercial indirect enzyme-linked immunosorbent assay. Anti-T. gondii antibodies were detected in 56 animals (13.3%, 95% CI: 10.1–16.6). A generalized estimating equations model revealed that the risk factors associated with higher seropositivity in pigs were altitude (higher in farm’s location < 250 m above sea level (masl) versus ≥ 250 masl) and age (higher in sows compared to post-weaning pigs). The results indicated that this protozoan parasite is widely distributed on pig farms in the study area, which is a public health concern since the consumption of raw or undercooked pork meat products containing tissue cysts is considered one of the main routes of T. gondii transmission worldwide. Control measures should be implemented to reduce the risk of exposure to T. gondii in pigs in Cuba.

scholarly journals An Integrated Study of Toxocara Infection in Honduran Children: Human Seroepidemiology and Environmental Contamination in a Coastal Community

2020 ◽  
Vol 5 (3) ◽  
pp. 135 ◽  
Author(s):  
Sergio A. Hernández ◽  
José A. Gabrie ◽  
Carol Anahelka Rodríguez ◽  
Gabriela Matamoros ◽  
María Mercedes Rueda ◽  
...  
Keyword(s):  
Logistic Regression ◽  
Soil Samples ◽  
Environmental Study ◽  
Enzyme Linked Immunosorbent Assay ◽  
Student Body ◽  
Nested Polymerase Chain Reaction ◽  
Serological Tests ◽  
Cross Sectional ◽  
Coastal Community ◽  
Pcr Rflp

(1) Background: Infections caused by Toxocara canis and T. cati are considered zoonoses of global importance. Reports from North and South America indicate that human infections are widespread in both continents, but epidemiological information from Central America is still lacking. (2) Methodology: In the present cross-sectional multi-year study, we aimed to undertake the first seroepidemiological and environmental study on toxocariasis in Honduras. This included the determination of seroprevalence of anti-Toxocara spp. antibodies in children using a Toxocara spp. purified excretory-secretory antigens enzyme-linked immunosorbent assay (TES-ELISA) and a confirmatory Western blot. As well, through statistical analysis including logistic regression we aimed at identifying relevant biological and epidemiological factors associated with seropositivity. The study also entailed detection of parasites’ eggs in the soil samples both through Sheather’s concentration method and a nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. (3) Results: The study was undertaken in a coastal community of Honduras in 2 different years, 2015 and 2017. A total of 88 healthy schoolchildren completed the study, with participation of 79% (73/92) and 65% (46/71) of the student body in 2015 and 2017, respectively. Thirty-one children participated in both years (i.e., dual participants). Through both serological tests, seropositivity was confirmed in 88.6% (78/88) of children. Due to the high number of seropositives, logistic regression analysis was not possible for most socio-economic and epidemiological variables. Eosinophilia, on the other hand, was associated with seropositivity, independently of other intestinal helminthic infections. Continued seropositivity was observed in most of the dual participants, while seroconversion was determined in 8 of these children. Microscopic examination of soil samples did not yield any positive results. Through nested PCR-RFLP, 3 of the 50 samples (6%) were positive for Toxocara spp.; two were identified as T. canis and one as T. cati. (4) Conclusions: This work documents for the first time, high levels of human exposure to Toxocara spp. in Honduras. These findings, along with the country’s favorable epidemiological conditions for this zoonosis, emphasize the need for more research to determine whether this infection is underreported in the country.

Evidence of the three main clonalToxoplasma gondiilineages from wild mammalian carnivores in the UK

Parasitology ◽  
2013 ◽  
Vol 140 (14) ◽  
pp. 1768-1776 ◽  
Author(s):  
A. BURRELLS ◽  
P. M. BARTLEY ◽  
I. A. ZIMMER ◽  
S. ROY ◽  
A. C. KITCHENER ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Rflp Markers ◽  
Type I ◽  
Type Ii ◽  
Red Foxes ◽  
Mammal Species ◽  
Tissue Samples ◽  
Pcr Rflp ◽  
Few Data ◽  
The Uk

SUMMARYToxoplasma gondiiis a zoonotic pathogen defined by three main clonal lineages (types I, II, III), of which type II is most common in Europe. Very few data exist on the prevalence and genotypes ofT. gondiiin the UK. Wildlife can act as sentinel species forT. gondiigenotypes present in the environment, which may subsequently be transmitted to livestock and humans. DNA was extracted from tissue samples of wild British carnivores, including 99 ferrets, 83 red foxes, 70 polecats, 65 mink, 64 badgers and 9 stoats. Parasite DNA was detected using a nested ITS1 PCR specific forT. gondii, PCR positive samples were subsequently genotyped using five PCR–RFLP markers.Toxoplasma gondiiDNA was detected within all these mammal species and prevalence varied from 6·0 to 44·4% depending on the host. PCR–RFLP genotyping identified type II as the predominant lineage, but type III and type I alleles were also identified. No atypical or mixed genotypes were identified within these animals. This study demonstrates the presence of alleles for all three clonal lineages with potential for transmission to cats and livestock. This is the first DNA-based study ofT. gondiiprevalence and genotypes across a broad range of wild British carnivores.

scholarly journals Potential linkage between Toxoplasma gondii Infection and physical education scores of College Students

2020 ◽  
Author(s):  
Zhijin Sheng ◽  
Yu Jin ◽  
Yinan Du ◽  
Xinlei Yan ◽  
Yong Yao
Keyword(s):  
College Students ◽  
Physical Education ◽  
Toxoplasma Gondii ◽  
Protozoan Parasite ◽  
Anhui Province ◽  
Behavioral Changes ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
School Students ◽  
Toxoplasma Gondii Infection

ABSTRACTObjectiveToxoplasma gondii is a worldwide protozoan parasite that could infect virtually all warm-blooded animals, including humans. Our study aimed to investigate the prevalence of T. gondii infection in college students at Anhui province, China. Moreover, growing studies demonstrated the association between T. gondii infection and host behavioral changes. We also studied the linkage between T. gondii and scores of college students.Methods2704 serum samples of medical school students attending physical education lessons were collected from September 2017 to September 2019 and evaluated for T. gondii IgG antibodies using an enzyme-linked immunosorbent assay (ELISA). We also analysed PE scores of T. gondii infected students and T. gondii uninfected students.ResultsThe overall seroprevalence of T. gondii was 11.5%. The main risk factors related to T. gondii infections were cat in the household and gardening or agriculture activity. Furthermore, in basketball group and football group, scores of T. gondii seropositive students were significantly higher than that of seronegative students, while in other sports there is no difference between scores of T. gondii infected students and T. gondii uninfected students.ConclusionThis is the first report of T. gondii seroprevalence in college students in Anhui province, China.

Sign in / Sign up

Export Citation Format

Share Document