Visceral larva migrans detection using PCR–RFLP in BALB/c mice infected with<i>Toxocara canis</i>

Özbakış, Gökben; Doganay, A.

doi:10.1017/s0022149x19000609

Cited by 7 publications

(7 citation statements)

References 36 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%

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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%

“…Interestingly, the more intense parasite DNA amplification bands coincided with the highest amount of parasite larvae in the tissues. A similar correlation of parasite specific DNA and amount of parasite larvae was described using a PCR–RFLP to evaluate parasite in tissue samples of BALB/c mice infected with T. canis [ 33 ].…”

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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“…It is worth mentioning that other Toxocara species infect humans, especially Toxocara cati . They share many molecules with T. canis and only the molecular diagnosis through PCR can distinguish among these infections (Özbakış & Doğanay, 2019). Because of the difficulty in differentiating the two species of Toxocara in the sera of infected individuals, we could not perform a cross‐reactivity assay using sera with antibodies against T. canis and T. cati .…”

Section: Discussionmentioning

confidence: 99%

Proteomics and immunoblotting analyses reveal antigens that optimize the immunodiagnosis of the infection by Toxocara spp

Silva

Fernandes

Silva

et al. 2022

Transbounding Emerging Dis

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Toxocariasis is an infection caused by the round worms Toxocara canis and Toxocara cati. It occurs worldwide though it is more prevalent in developing countries. For the diagnosis of toxocariasis, the most used method is the indirect enzyme-linked immunosorbent assay (indirect ELISA), based on the detection of specific antibodies using the excreted/secreted products from T. canis larvae (TES) as antigens, but it crossreacts with several helminth infections. For this reason, there is a need to investigate species-specific immunoreactive proteins, which can be used for the development of a more sensitive and specific diagnosis. This study aims to investigate immunoreactive protein candidates to be used for the development of a more sensitive and specific diagnosis of Toxocara spp. infection in humans. We have used immunoblotting and mass spectrometry to select four Toxocara canis immunoreactive proteins that were recombinantly expressed in bacteria and evaluated as potential new diagnostic antigens (rMUC3, rTES 26, rTES32 and rCTL4). The recognition of these recombinant proteins by total serum IgG and IgG4 was assayed using the purified proteins in an isolated manner or in combination. The IgG ELISAs performed with individual recombinant antigens reached values of sensitivity and specificity that ranged from 91.7% to 97.3% and 94.0% to 97.9%, respectively. Among the analyses, the IgG4 immunoassay was proven to be more effective, revealing a sensitivity that ranged from 88.8% to 98.3% and a specificity of 97.8%-97.9%. The IgG4 ELISA was shown to be more effective and presented no cross-reactivity when using combinations of the rTES 26 and rCTL4 recombinant proteins. The combination of these two molecules achieved 100% sensitivity and specificity. The use of only two recombinant proteins can contribute to improve the current panorama of toxocariasis immunodiagnosis for, with a better optimization and reduced cost.

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“…Another study reported increased levels of serum IL-6 in mice infected with T. canis eggs compared with the control group [34]. Other surveys showed the use of a molecular method (PCR–RFLP) to diagnose toxocariasis in BALB/c mice [35]. Another study compared the prevalence of toxocariasis in dogs using ELISA (IgG antibody) and PCR (based on ITS1 rRNA) and found more positive cases from the serological test (71.0 %) compared with the molecular method (58.0 %) [36].…”

Section: Discussionmentioning

confidence: 99%

Changes in the expression of miR-103a and miR-21: a functional diagnosis of toxocariasis in rats

Zibaei

Rokni

Ibrahim

et al. 2022

Journal of Medical Microbiology

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Introduction. Toxocariasis is a zoonotic parasitic disease caused by migrating nematode worms, Toxocara species larvae, within tissues. MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression at a post-transcriptional level. Hypothesis/Gap Statement. miRNA-based diagnostic biomarkers for toxocariasis are emerging, but there is limited information about the role of many miRNAs and a more detailed diagnostic evaluation of miRNA expression patterns is needed to understand their immunobiological function. Aim. We investigated the expression levels of circulating miRNA 21 and miRNA 103a as potential biomarkers for the prediction and diagnosis of toxocariasis in Wistar rats infected with Toxocara canis. Methodology. Thirty Wistar rats were inoculated orally with 2500 T. canis embryonated eggs via gavage. Serum samples were collected from infected animals and were tested against T. canis antigens for 60 days post-infection. The plasma samples were isolated for quantitative real-time PCR (qPCR) assays and qPCR was used to assess transcription levels of miRNA 21 and miRNA 103a. Results. The prevalence of anti-Toxocara IgG was detected in 7/30 (23.3 %) infected rats. Molecular analysis of miRNAs 21 and 103a showed that expression levels of miRNAs in both groups of Toxocara-positive and negative samples were the same without significant association. The ratio of housekeeping gene expression (U6) to gene expression of miRNAs 21 and 103a indicated the rate of change (1/1.38 ≈ 0.75 and 1/0.751 ≈ 1.3, respectively). Conclusion. Our study revealed that miRNAs 21 and 103a might play fundamental roles as biomarkers and diagnostic tools for toxocariasis. However, the changes in expression of these miRNAs were not adequate to be used as biomarkers in diagnosis.

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

Cited by 7 publications

References 36 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

Proteomics and immunoblotting analyses reveal antigens that optimize the immunodiagnosis of the infection by Toxocara spp

Changes in the expression of miR-103a and miR-21: a functional diagnosis of toxocariasis in rats

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