DNA Fingerprinting of Cryptosporidium parvum Isolates Using Amplified Fragment Length Polymorphism (AFLP)

Blears, M.; Pokorny, Nicholas J.; Carreno, Ramon A.; Chen, Shu; Grandis, Stephanie A. De; Lee, Hung; Trevors, J. T.

doi:10.1645/0022-3395(2000)086[0838:dfocpi]2.0.co;2

Cited by 21 publications

(7 citation statements)

References 17 publications

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“…Isolates WH33 and WH37 were identical to Cryptosporidium genotype W17 deposited in GenBank (GenBank accession number AY737573 [13]). WH3 was identical to cervine genotype sequences (GenBank accession numbers AY737592 [13], AJ849465, and AF442484 [4]), while WH5 was a variant of the cervine genotype that differed by an AT deletion at positions 689 and 690 of the C. parvum 18S rRNA gene (GenBank accession number AF164102 [2]) (Fig. 2).…”

Section: Resultsmentioning

confidence: 97%

Evidence Supporting Zoonotic Transmission of Cryptosporidium spp. in Wisconsin

et al. 2006

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Cryptosporidium hominis and Cryptosporidium parvum are the primary species of Cryptosporidium that infect humans. C. hominis has an anthroponotic transmission cycle, while C. parvum is zoonotic, infecting cattle and other ruminants, in addition to humans. Most cryptosporidiosis outbreaks in the United States have been caused by C. hominis, and this species is often reported as the primary cause of cryptosporidiosis in this country. However, outbreaks account for only 10% of the overall cryptosporidiosis cases, and there are few data on the species that cause sporadic cases. The present study identified the species/genotypes and subgenotypes of Cryptosporidium in 49 cases of sporadic cryptosporidiosis in Wisconsin during the period from 2003 to 2005. The species/genotype of isolates was determined by PCR restriction fragment length polymorphism analysis of the 18S rRNA and Cryptosporidium oocyst wall protein genes. The C. parvum and C. hominis isolates were subgenotyped by sequence analysis of the GP60 gene. Forty-four of 49 isolates were identified as C. parvum, and 1 was identified as C. hominis. Of the remaining isolates, one was identified as being of the cervine genotype, one was identified as being a cervine genotype variant, and two were identified as being of a novel human genotype, previously reported as W17. Nine different subgenotypes were identified within the C. parvum species, and two of these were responsible for 60% of the cases. In this study we found that most sporadic cases of cryptosporidiosis in Wisconsin are caused by zoonotic Cryptosporidium species, indicating that zoonotic transmission could be more frequently associated with sporadic cases in the United States.

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Section: Resultsmentioning

confidence: 97%

Evidence Supporting Zoonotic Transmission of Cryptosporidium spp. in Wisconsin

et al. 2006

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“…It consists of four steps: DNA digestion, ligation, amplification, and gel analysis. Polymorphisms are identified based on the presence or absence of DNA fragments by polyacrylamide gel analysis (7). The advantages of this technique are the ability to search an entire genome for polymorphisms, the reproducibility of the method, and the possibility of being used against parasites about which there is no prior genetic information (7,77,78).…”

Section: Amplified Fragment Length Polymorphism (Aflp)mentioning

confidence: 99%

“…Studies have demonstrated its application to differentiate isolates of C. parvum into two distinct genotypes, as well as strains of Leishmania belonging to cutaneous leishmaniosis (CL) and visceral leishmaniosis (VL) (7). Analysis based on AFLP polymorphic markers have revealed high genetic variability among the genome species of Leishmania major, L. tropica and L. donovani, which was sufficient to distinguish between CL and VL (79).…”

Section: Amplified Fragment Length Polymorphism (Aflp)mentioning

confidence: 99%

“…Current laboratory diagnostic methods for the identification of parasites include: polymerase chain reaction (PCR), random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), microsatellite marker method, Luminex xMAP-based technology (areas of multianalyte profiling), loop-mediated isothermal amplification (LAMP), and the recently added real-time PCR (5)(6)(7)(8)(9)(10)(11)(12).…”

Section: Introductionmentioning

confidence: 99%

“…Polymorphisms are identified based on the presence or absence of DNA fragments by polyacrylamide gel analysis (7). The advantages of this technique are the ability to search an entire genome for polymorphisms, the reproducibility of the method, and the possibility of being used against parasites about which there is no prior genetic information (7,77,78).Studies have demonstrated its application to differentiate isolates of C. parvum into two distinct genotypes, as well as strains of Leishmania belonging to cutaneous leishmaniosis (CL) and visceral leishmaniosis (VL) (7). Analysis based on AFLP polymorphic markers have revealed high genetic variability among the genome species of Leishmania major, L. tropica and L. donovani, which was sufficient to distinguish between CL and VL (79).…”

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Molecular techniques for the study and diagnosis of parasite infection

Tavares¹,

Staggemeier²,

Borges³

et al. 2011

J. Venom. Anim. Toxins incl. Trop. Dis

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Abstract:In parasitology, routine laboratory diagnosis involves conventional methods, such as optical microscopy, used for the morphological identification of parasites. Currently, molecular biology techniques are increasingly used to diagnose parasite structures in order to enhance the identification and characterization of parasites. The objective of the present study was to review the main current and new diagnostic techniques for confirmation of parasite infections, namely: polymerase chain reaction (PCR), real-time polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), Luminex xMAP, random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and restriction fragment length polymorphism (RFLP), in addition to microsatellites. Molecular assays have comprehensively assisted in the diagnosis, treatment and epidemiological studies of parasitic diseases that affect people worldwide, helping to control parasitic disease mortality.Key words: parasite infection, diagnosis, molecular techniques, molecular epidemiology. Review ARticle The Journal of Venomous Animals and Toxins including Tropical Diseases ISSN 1678-9199 | 2011 | volume 17 | issue 3 | pages 239-248 INTRODUCTIONIn parasitology, routine laboratory diagnosis involves conventional methods, such as optical microscopy, used for the morphological identification of parasites (1). However, the occasional difficulty of identifying these parasite structures may decrease the sensitivity of such methods. Currently, because of these difficulties, molecular biology has been employed to detect parasites responsible for parasitic diseases (2).Traditional parasitological analyses have the advantage of being less costly without requiring expensive reagents and equipment. Additionally, such analyses can be easily performed when a trained microscopist is available. On the other hand, molecular technology demonstrates the presence of parasites based on their antigenic components or DNA segments (3). These tests are not influenced by environmental factors that usually can interfere with the results of a stool test, for example, thus ensuring highly reliable results (4).Current laboratory diagnostic methods for the identification of parasites include: polymerase chain reaction (PCR), random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), microsatellite marker method, Luminex xMAP-based technology (areas of multianalyte profiling), loop-mediated isothermal amplification (LAMP), and the recently added real-time PCR (5-12).The objective of the present article was to review the applications of molecular biology techniques in parasitology by evaluating and discussing their uses and benefits. MOLECULAR METHODS FOR DETECTION OF PARASITE STRUCTURESSeveral molecular tests to detect parasites have been developed in the last decade. Their specificity and sensitivity have gradually increased, and parasites that were previously difficult to diagnose usi...

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Molecular Characterization and Taxonomy of Cryptosporidium

Ryan¹

2003

Cryptosporidium

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DNA Fingerprinting of Cryptosporidium parvum Isolates Using Amplified Fragment Length Polymorphism (AFLP)

Cited by 21 publications

References 17 publications

Evidence Supporting Zoonotic Transmission of Cryptosporidium spp. in Wisconsin

Evidence Supporting Zoonotic Transmission of Cryptosporidium spp. in Wisconsin

Molecular techniques for the study and diagnosis of parasite infection

Molecular Characterization and Taxonomy of Cryptosporidium

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