Development of nested multiplex polymerase chain reaction (PCR) assay for the detection of Toxocara canis , Toxocara cati and Ascaris suum contamination in meat and organ meats

Wang, Zhenzhen; Shibata, Miho; Nguyen, Yen Thi Hoang; Hayata, Yayoi; Nonaka, Nariaki; Maruyama, Haruhiko; Yoshida, Ayako

doi:10.1016/j.parint.2018.06.006

Cited by 16 publications

(10 citation statements)

References 30 publications

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“…If migrating, their presence in tissue samples can be detected by PCR. The fact that PCR was positive throughout the recovery period while the microscopic method was only positive for inoculation at the beginning indicates that PCR assay is a better choice when the prevalence of infection or intensity of infection is low [ 32 , 33 ]. Interestingly, the more intense parasite DNA amplification bands coincided with the highest amount of parasite larvae in the tissues.…”

Section: Discussionmentioning

confidence: 99%

Larva migrans in BALB/c mice experimentally infected with Toxocara cati ensured by PCR assay

et al. 2022

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Background Toxocara cati, the cat roundworm, is a parasitic nematode that known to cause toxocariasis in intermediate hosts and humans. In this study, we characterized the dynamics of T. cati larvae migration in BALB/c mice after inoculation with eggs and ensured the migration detecting the larval DNA by a PCR. To evaluate the dynamics of larval migration and distribution, twenty-four BALB/c mice were orally inoculated with 2500 T. cati infective eggs and the visceral organs of the infected animals were examined by pepsin digestion and microscopic parasite counts, followed by PCR at day 1 to 28 post-inoculation. Results The PCR assays were successfully used for detection of T. cati larvae in tissue samples and T. cati larvae and the DNAs were found in the liver, lungs, heart, kidneys and the brain. We detected T. cati in 92.2% of tissue samples by PCR, 30% higher than the conventional pepsin digestion technique. Conclusion Our findings demonstrated that the PCR assay is a sensitive and specific for the detection of T. cati larvae. Therefore, it could become a useful tool for the investigation of the dynamics of larval migration and Toxocara infection in murine model.

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Section: Discussionmentioning

confidence: 99%

Larva migrans in BALB/c mice experimentally infected with Toxocara cati ensured by PCR assay

et al. 2022

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“…Some studies were performed with different amounts of tissues to isolate DNA (Wang et al ., 2018). According to our study design, we used 20 mg of tissue and obtained positive PCR results.…”

Section: Discussionmentioning

confidence: 99%

“…The organs (liver, heart, lung, kidney, brain and intestine) of paratenic hosts were investigated with the digestion method and PCR technique. The results of both techniques revealed that PCR is more reliable and able to detect species of parasites (Zibaei et al ., 2017; Wang et al ., 2018). Although larvae could be observed by using the digestion method, this approach took a long time and required extra effort.…”

Section: Discussionmentioning

confidence: 99%

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Visceral larva migrans detection using PCR–RFLP in BALB/c mice infected withToxocara canis

Özbakış

Doganay

2019

J. Helminthol.

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Toxocara canis is an important zoonotic roundworm distributed worldwide. The infective larvae of T. canis are one of the causes of visceral larva migrans (VLM), a clinical syndrome in humans. Diagnosing VLM is difficult, and the differential diagnosis of the larval development stage is limited. Therefore, this experimental research aimed to diagnose T. canis larvae using a molecular method, not only in liver tissue, which is the most commonly affected tissue, but also in the limb muscles, lungs and brain tissues. For this purpose, 24 BALB/c mice were infected with 1000 embryonated T. canis eggs. Necropsies were performed on the second, fourth, seventh and 14th days post-infection. While a part of the samples were digested with pepsin-HCl, the molecular method was used for the remainder of the samples to replicate the mitochondrial DNA adenosine triphosphate (ATP) synthase subunit-6 gene region of T. canis. BbsI, a restriction endonuclease, was used to determine the specificity of the amplicons obtained from Polymerase chain reaction (PCR). The detection limit for embryonated eggs was recorded. The PCR results showed that the sensitivity of the PCR analysis was 83.3% in the liver (with 88.8% accuracy), 87.5% in the lungs (with 91.6% accuracy) and 75.0% in the brain, forelimb and hindlimb muscles (with 83.3% accuracy). In all tissues, the test specificity was determined to be 100%. In this study, the molecular method was applied to only experimentally infected BALB/c mice tissues; thus, it is suggested that it can be also employed in different paratenic hosts and materials possibly infected with T. canis.

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“…Indeed, Toxocara larvae are frequently seen as a 'contaminant' when isolated during routine porcine Trichinella testing by artificial tissue digestion (Marucci et al, 2013), and their presence is often not reported. Some recent studies have advocated the use of molecular methods to improve the detection of helminth contamination of meat and offal (Nguyen et al, 2017;Wang et al, 2018;Karadjian et al, 2020). This is an area which has the potential to greatly improve the detection of Toxocara during the meat inspection process and warrants further investigation.…”

Section: Meat Inspection At Slaughtermentioning

confidence: 99%

Brain food: rethinking food-borne toxocariasis

et al. 2021

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Human toxocariasis is a neglected tropical disease, which is actually global in distribution and has a significant impact on global public health. The infection can lead to several serious conditions in humans, including allergic, ophthalmic and neurological disorders such as epilepsy. It is caused by the common roundworm species Toxocara canis and Toxocara cati, with humans becoming accidentally infected via the ingestion of eggs or larvae. Toxocara eggs are deposited on the ground when infected dogs, cats and foxes defecate, with the eggs contaminating crops, grazing pastures, and subsequently food animals. However, transmission of Toxocara to humans via food consumption has received relatively little attention in the literature. To establish the risks that contaminated food poses to the public, a renewed research focus is required. This review discusses what is currently known about food-borne Toxocara transmission, highlighting the gaps in our understanding that require further attention, and outlining some potential preventative strategies which could be employed to safeguard consumer health.

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Development of nested multiplex polymerase chain reaction (PCR) assay for the detection of Toxocara canis , Toxocara cati and Ascaris suum contamination in meat and organ meats

Cited by 16 publications

References 30 publications

Larva migrans in BALB/c mice experimentally infected with Toxocara cati ensured by PCR assay

Larva migrans in BALB/c mice experimentally infected with Toxocara cati ensured by PCR assay

Visceral larva migrans detection using PCR–RFLP in BALB/c mice infected withToxocara canis

Brain food: rethinking food-borne toxocariasis

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