Rapid Microsphere Assay for Identification of
            <i>Cryptosporidium hominis</i>
            and
            <i>Cryptosporidium parvum</i>
            in Stool and Environmental Samples

Bandyopadhyay, Kaushik; Kellar, Kathryn L.; Moura, Iaci N. S.; Carollo, Maria Cristina Casaqui; Graczyk, Thaddeus K.; Slemenda, Susan B.; Johnston, Stephanie; Silva, Alexandre J. da

doi:10.1128/jcm.00138-07

Cited by 31 publications

(30 citation statements)

References 39 publications

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“…Luminex beadarrays have been developed for detecting microbial pathogens from clinical (e.g. Bandyopadhyay et al 2007) and environmental (Baums et al 2007) samples. Applications of this technology have also been used to determine bacteria and phytoplankton abundances from the marine environment (Ellison & Burton 2005, Scorzetti et al 2009, Mayali et al 2010.…”

mentioning

confidence: 99%

Temporal and spatial distributions of marine Synechococcus in the Southern California Bight assessed by hybridization to bead-arrays

Tai¹,

Burton²,

Palenik

2011

Mar. Ecol. Prog. Ser.

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Marine Synechococcus diversity has been previously described using multi-locus gene sequence phylogenies and the identification of distinct clades. Synechococcus from Clades I, II, III, and IV and from sub-clades within Clades I and IV were enumerated from environmental samples by developing a hybridization assay to liquid bead-arrays (Luminex). Oligonucleotide probes targeting a gene encoding a subunit of RNA polymerase (rpoC1) were used simultaneously in multiplexed assays to track Synechococcus diversity from a Pacific Ocean coastal monitoring site and along a coastal to open-ocean transect in the Southern California Bight. The Luminex assay demonstrated that Synechococcus from Clades I and IV were the dominant types at the coastal site throughout the year. Synechococcus from Clades II and III were not detected except during the late summer or early winter. Within the dominant Clades I and IV, rpoC1-defined sub-clades of Synechococcus showed distinct spatial distributions along the coastal to open-ocean transect, coinciding with changes in the nitricline, thermocline, and fluorescence (chlorophyll) maximum depths. In coastal waters, Synechococcus targeted by 2 sub-clade IV probes were dominant at the surface, whereas 2 sub-clade I probes and a third sub-clade IV probe had increased signals in deeper water near the fluorescence maximum. In mesotrophic waters, this third sub-clade IV probe dominated at the fluorescence maximum (depth of 50 to 70 m), whereas all other sub-clade probes were below detection limits. The differing distributions of sub-clades within the dominant Synechococcus clades indicate that the subclades likely have adapted to distinct ecological niches found within the Southern California Bight. KEY WORDS: Cyanobacteria · Microbial ecology · Biogeography · Time-series · LuminexResale or republication not permitted without written consent of the publisher Mar Ecol Prog Ser 426: 133-147, 2011, Zwirglmaier et al. 2008) and off-shore oligotrophic environments (Ferris & Palenik 1998, Toledo & Palenik 2003, Ahlgren & Rocap 2006. Physiologically, differences in growth rates, in nitrogen and phosphate utilization, in pigment composition, in chromatic adaptation, and in motility have typically been used to describe isolated strains of Synechococcus (Moore et al. 1995, Collier et al. 1999, Toledo et al. 1999, Ahlgren & Rocap 2006, but, with the exception of motility (which has been found only in Clade III isolates), phenotypic traits and adaptations that uniquely characterize the genetically defined clades have yet to be identified.To understand the ecology of Synechococcus, diversity and abundance measures are fundamental. However, due to their small size, obtaining these measures is technically challenging. Traditionally, microbes are often distinguished physiologically by studying cultured isolates. But since only a small fraction of microbial diversity is represented by cultured isolates, and since the process of cultivation is labor-intensive, culture-independent and molecular methods h...

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mentioning

confidence: 99%

Temporal and spatial distributions of marine Synechococcus in the Southern California Bight assessed by hybridization to bead-arrays

Tai¹,

Burton²,

Palenik

2011

Mar. Ecol. Prog. Ser.

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“…Of the two detection reagents added, one decays to hydrogen peroxide and is reduced by the enzyme peroxidase. The reduction reaction causes the luminol in the second detection reagent to be oxidized (Kong & Gilbert, 2007;Bandyopadhyay et al, 2007). The resulting by-products emit light, which is detected by sensitive photographic film or a compact flow cytometer.…”

Section: Reverse Line Hybridization (Rlh)mentioning

confidence: 99%

“…Continual improvements of RLH have resulted in a new method that eliminates the DNA sequencing step after PCR (Bandyopadhyay et al, 2007). This so-called Luminex-based assay can differentiate between C. hominis and C. parvum in a rapid and cost-effective manner.…”

Section: Luminex-based Assay As An Improved Version Of Rlhmentioning

confidence: 99%

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Potential molecular tools for assessing the public health risk associated with waterborne Cryptosporidium oocysts

Kothavade¹

2012

Journal of Medical Microbiology

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“…Luminex was applied to the study of Cryptosporidium by Bandyopadhyay et al (64). The species C. hominis and C. parvum differ genetically by a single nucleotide in the microsatellite region-2 (ML-2), which cannot be distinguished using antigen detection or serology tests.…”

Section: Luminexmentioning

confidence: 99%

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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Rapid Microsphere Assay for Identification of Cryptosporidium hominis and Cryptosporidium parvum in Stool and Environmental Samples

Cited by 31 publications

References 39 publications

Temporal and spatial distributions of marine Synechococcus in the Southern California Bight assessed by hybridization to bead-arrays

Temporal and spatial distributions of marine Synechococcus in the Southern California Bight assessed by hybridization to bead-arrays

Potential molecular tools for assessing the public health risk associated with waterborne Cryptosporidium oocysts

Molecular techniques for the study and diagnosis of parasite infection

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