Detection and identification of Giardia species using real-time PCR and sequencing

Jothikumar, Narayanan; Murphy, Jennifer L.; Hill, Vincent R.

doi:10.1016/j.mimet.2021.106279

Cited by 7 publications

(3 citation statements)

References 26 publications

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“…The qPCR is widely used in the detection of pathogens due to its great sensitivity and savings in time and effort, in addition there are different variants for this technique such as Molecular Beacon probes, Taqman probes, Scorpion probes, FRET probes, and intercalating dyes such as SYBR Green, that alone or in combination with other techniques have been used for the detection, quantification, and characterization of Giardia [ 92 , 106 , 107 , 108 , 109 ].…”

Section: Discussionmentioning

confidence: 99%

Burden and Epidemiology of Human Intestinal Giardia duodenalis Infection in Colombia: A Systematic Review

et al. 2022

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The genus Giardia is a unicellular protozoan able to parasitize both humans and animals. Cysts of Giardia can be found in soil samples, aquatic environments, food, and any surface that gets in contact with the feces of parasitized animals. The aim of this systematic review was to analyze the burden and epidemiology of Giardia infection in Colombia summarizing recent scientific reports and existing knowledge and to identify knowledge gaps that may be addressed in future investigations. This work follows the guidelines established by “Preferred Reporting Items for Systematic Reviews and Meta-Analyzes” (PRISMA). Published scientific literature from 1 January 2010 to 18 September 2022 was searched in six electronic scientific databases using the search terms: “Giardia” OR “Giardiasis” AND “Colombia”. Twenty-three scientific articles were performed in 22 departments of Colombia at rural, urban, and a combination of rural and urban contexts. The prevalence of Giardia in the Colombian population was between 0.9 and 48.1% when the samples were analyzed with classical microscopy; the range of Giardia prevalence was even bigger (4.2–100%) when qPCR and nested PCR were used. The dominant Giardia assemblages found in Colombia were A and B, and most frequent subassemblages were AII, BIII, and BIV.

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Section: Discussionmentioning

confidence: 99%

Burden and Epidemiology of Human Intestinal Giardia duodenalis Infection in Colombia: A Systematic Review

et al. 2022

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“…Real-time polymerase chain reaction (PCR) with a hydrolysis probe was performed for the detection of Salmonella ( ttr gene), Cryptosporidium (18S rRNA), Giardia (18S rDNA), Naegleria fowleri (ITS), pathogenic Leptospira ( LipL32 gene), and a marker of human fecal contamination, Bacteroides HF183 (HF183) (Table S1). − Although HF183 is not a pathogen, this human-specific fecal marker is useful in determining the prevalence of human fecal contamination that could contain enteric pathogens in cisterns (e.g., via sewage or septic intrusion). Five microliters of template DNA were analyzed in triplicate in 50 μL PCR reaction volumes for all assays except Leptospira , which used 20 μL reaction volumes.…”

Section: Methodsmentioning

confidence: 99%

Microbial Characterization, Factors Contributing to Contamination, and Household Use of Cistern Water, U.S. Virgin Islands

et al. 2022

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Households in the United States Virgin Islands (USVI) heavily rely on roof-harvested rainwater stored in cisterns for their daily activities. However, there are insufficient data on cistern water microbiological and physicochemical characteristics to inform appropriate cistern water management. Cistern and kitchen tap water samples were collected from 399 geographically representative households across St. Croix, St. Thomas, and St. John and an administered survey captured household site and cistern characteristics and water use behaviors. Water samples were analyzed for Escherichia coli by culture, and a subset of cistern water samples (N = 47) were analyzed for Salmonella, Naegleria fowleri, pathogenic Leptospira, Cryptosporidium, Giardia, and human-specific fecal contamination using real-time polymerase chain reaction (PCR). Associations between E. coli cistern contamination and cistern and site characteristics were evaluated to better understand possible mechanisms of contamination. E. coli was detected in 80% of cistern water samples and in 58% of kitchen tap samples. For the subset of samples tested by PCR, at least one of the pathogens was detected in 66% of cisterns. Our results suggest that covering overflow pipes with screens, decreasing animal presence at the household, and preventing animals or insects from entering the cisterns can decrease the likelihood of E. coli contamination in USVI cistern water.

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“…flotation or sedimentation), but their predictive value may be affected by the intermittent shedding of cysts in the host excrements and the expertise of the diagnostician [ 15 ]. Molecular methods, such as conventional PCR (cPCR) and real-time PCR (qPCR) [ 15 , 16 ], are also used for detection and assemblage identification. However, the identification of assemblages is impeded by the limited sensitivity of existing molecular genotyping tools, particularly in individuals with weak colonization [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

The opportunistic protist, Giardia intestinalis , occurs in gut-healthy humans in a high-income country

Brožová,

Jirků,

Lhotská

et al. 2023

Emerging Microbes & Infections

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Giardia intestinalis , a cosmopolitan gastrointestinal protist, is detected mainly in patients with clinical giardiasis in high-income countries. In contrast, there is very little information on the presence of Giardia in asymptomatic individuals. Therefore, the aim of this study was to determine the presence and prevalence of Giardia in gut-healthy volunteers in the Czech Republic and to perform a comparative evaluation of different diagnostic methods, since Giardia diagnostics is complicated. Our results confirmed that the qPCR method is the most sensitive method for detecting Giardia and revealed a prevalence of 7% (22/296) in asymptomatic individuals. In most cases, the colonization intensity ranged from 10 −1 –10 1 . A conventional PCR protocol targeting the TPI gene was used to identify the assemblages. However, this protocol had limited sensitivity for Giardia amplification, effectively detecting colonization above an intensity of 10 4 . In addition, Giardia was detected in 19% of the animals, which were closely associated with the study participants. However, due to methodological limitations, zoonotic transmission could not be clearly confirmed. Notably, contact with animals proved to be the only factor that had a significant impact on the incidence of Giardia in gut-healthy humans.

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Detection and identification of Giardia species using real-time PCR and sequencing

Cited by 7 publications

References 26 publications

Burden and Epidemiology of Human Intestinal Giardia duodenalis Infection in Colombia: A Systematic Review

Burden and Epidemiology of Human Intestinal Giardia duodenalis Infection in Colombia: A Systematic Review

Microbial Characterization, Factors Contributing to Contamination, and Household Use of Cistern Water, U.S. Virgin Islands

The opportunistic protist, Giardia intestinalis , occurs in gut-healthy humans in a high-income country

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