Toxoplasma gondii: Sensitive and rapid detection of infection by loop-mediated isothermal amplification (LAMP) method

Zhang, Houshuang; Thekisoe, Oriel; Aboge, Gabriel Oluga; Kyan, Hisako; Yamagishi, Junya; Inoue, Noboru; Nishikawa, Yoshifumi; Zakimi, Satoshi; Xuan, Xuenan

doi:10.1016/j.exppara.2009.01.012

Cited by 80 publications

(62 citation statements)

References 26 publications

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“…Several reports were confirmed the higher sensitivity of LAMP for detection of T. gondii than conventional PCR targeting 529 bp fragment of T. gondii genome (Fallahi et al 2014;Kong et al 2012;Lin et al 2012;Zhang et al 2009) or other specific targeting of T. gondii DNA regions (Fallahi et al 2014;Hu et al 2012;Kong et al 2012;Krasteva et al 2009;Lau et al 2010;Lin et al 2012;Sotiriadou and Karanis 2008;Zhang et al 2009). Until now, LAMP method was used for detection of T. gondii in pig (Lili et al 2014;Lin et al 2012;Qu et al 2013;Wang et al 2013;Zhang et al 2009), sheep (Lin et al 2012), mice (Hu et al 2012;Kong et al 2012;Krasteva et al 2009), and human (Fallahi et al 2014;Lau et al 2010). LAMP was also used for detection of T. gondii oocysts in water samples (GallasLindemann et al 2013;Koloren and Demirel 2013;Sotiriadou and Karanis 2008).…”

Section: Discussionmentioning

confidence: 80%

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Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

et al. 2015

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Toxoplasma gondii is one of the most common zoonotic parasitic diseases in human and warm-blooded animals worldwide. Birds are one of important intermediate hosts of T. gondii. The aim of this study is molecular detection of T. gondii in the house sparrow by LAMP and PCR methods in Tehran, Iran. A total 200 sparrows were captured in different regions of Tehran. DNA was extracted from tissue samples of each sparrow. LAMP and conventional PCR assays were carried out with a set of primers to detect the 529 bp fragment of T. gondii. LAMP and PCR were detected T. gondii from 17 (8.5 %) and 15 (7.5 %) of 200 sparrows respectively. These results indicated that sensitivity of LAMP was higher than conventional PCR. In our knowledge, this study is the first report of detection of T. gondii by LAMP method in bird hosts. Also, these findings provided an insight into epidemiological pattern of T. gondii infection in sparrow in Iran.

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Section: Discussionmentioning

confidence: 80%

“…LAMP has been developed for molecular detection of several microorganisms [reviewed in (Mori et al 2013)], including T. gondii (Kong et al 2012;Lin et al 2012;Zhang et al 2009). …”

Section: Introductionmentioning

confidence: 99%

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

et al. 2015

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“…[24] The cost of PCR is high and its application under field conditions is limited by the need for a thermocycler. Alternative amplification of DNA by loop-mediated isothermal amplification, using the 529 element, has been determined [10,25] and should be developed further for application in point-of-care diagnosis.…”

Section: Discussionmentioning

confidence: 99%

Use of the nested polymerase chain reaction for detection of Toxoplasma gondii in slaughterhouse workers in Thika District, Kenya

Thiongo¹,

Ichagichu

Ngotho

et al. 2016

S Afr Med J

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Background. The widely used methods of diagnosis of Toxoplasma gondii are serological. Current reports indicate a high seroprevalence of T. gondii in humans in Kenya. There is a need for more sensitive diagnostic tests, especially when the specific antibody titres are below detectable threshold levels. Use of the polymerase chain reaction (PCR) targeting the repetitive 529 base pair loci has been reported to be sensitive and specific. Objective. To detect T. gondii in a high-risk group of public health workers in Thika District, Kenya. Methods. In total, 87 human blood samples were collected from male slaughterhouse workers between 1 March 2013 and 25 June 2013. The DNA extracted was amplified by the nested PCR. Results. T. gondii was detected in 39.1% (34/87) of the workers. In the cow-sheep-goat slaughterhouses the prevalence ranged between 20% and 60%, while all the chicken slaughterhouse workers (6/6, 100%) tested positive. The difference in T. gondii positivity between the workers in the chicken slaughterhouse and those in the cattle-sheep-goat slaughterhouses was statistically significant (p=0.003). Conclusion. This study shows the presence of T. gondii in an asymptomatic high-risk group in Thika District, indicating the need for enhancement of public health awareness.

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“…This technique is more sensitive than conventional PCR but low sensitive than real time PCR [53]. The LAMP assay has been evaluated identifying REP 529-bp, SAG1, SAG2 and B1 T. gondii sequences from PBMC [54]. The reported sensitivity was 0.1 tachyzoites and crossed reactivity with other parasites was not found.…”

Section: Loop Mediated Isothermal Ampliication (Lamp)mentioning

confidence: 99%

“…The inal products are multiple loop DNA formed by alignment between inverted repeated sequences present in the same chain [53]. It only requires a water bath or thermo block to realize the ampliication of DNA and the ampliied products can be analysed by real time PCR equipment or by electrophoresis [54]. This technique is more sensitive than conventional PCR but low sensitive than real time PCR [53].…”

Section: Loop Mediated Isothermal Ampliication (Lamp)mentioning

confidence: 99%

The Laboratory Diagnosis in Toxoplasma Infection

Ramírez¹,

Orozco²,

Ramírez³

2017

Toxoplasmosis

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The diagnosis of toxoplasmosis is of great importance due to the damage caused by this parasite in immunosuppressed people or in pregnant women, the diagnosis of an active toxoplasmosis represents a sign to initiate a pharmacological treatment immediately. The diagnosis of Toxoplasma can be performed with direct methods through intraperitoneal inoculation of serum or cerebrospinal luid, in susceptible mice evaluating the survival and detection of tachyzoites of biological samples. Indirect methods detecting the IgM and IgG isotypes against Toxoplasma have been the tools mostly used and had leaded to discriminate between an active and acute, from a chronic toxoplasmosis. Molecular methods actually Toxoplasma-DNA identiication by molecular biology tests like the polymerase chain reaction (PCR) allow the direct detection of the parasite. Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) have been used to identify three strain linages (type I, II and III). Recently, a high-resolution melting method was described to determine the genotype of the infection by Toxoplasma gondii directly from biological samples.

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Toxoplasma gondii: Sensitive and rapid detection of infection by loop-mediated isothermal amplification (LAMP) method

Cited by 80 publications

References 26 publications

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

Use of the nested polymerase chain reaction for detection of Toxoplasma gondii in slaughterhouse workers in Thika District, Kenya

The Laboratory Diagnosis in Toxoplasma Infection

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