The use of ELISA, nPCR and qPCR for diagnosis of ocular toxoplasmosis in experimentally infected pigs
Copyright © 2017 Elsevier Ltd. All rights reserved..
In the present study we experimentally infected pigs with T. gondii tachyzoites, bradyzoites and oocysts in order to evaluate IgG-ELISA, nested-PCR, and qPCR for diagnosis of ocular infection. Eighteen pigs were divided into four groups: G1 (infected with 103 tissue cysts of the M4 strain (type II) at day 28, n=5), G2 (infected with 103 oocysts of the M4 strain at day 28, n=5), G3 (infected with tachyzoites of S48 strain (type 1) at day 0, n=5), and G4 (uninfected unchallenged, control group n=3). At day 70 of the experiment all animals were culled, and serum, aqueous humor (AH) and vitreous humor (VH) samples were collected to perform indirect ELISA, and PCR (nPCR, and qPCR). By ELISA nine pigs (60%) out of 15 were positive in VH samples, and seven out of 15 (46%) were positive in AH samples. Both molecular techniques used here, nPCR and qPCR, were able to detect <50fg of T. gondii tachyzoite DNA. The nPCR and qPCR detected six (7/15, 47%) and two (2/15, 13.3%) positive animals respectively. Antibody responses were detected in serum and in AH and VH from the eye, suggesting that pigs may be an animal that could be used as a model to further our understanding of diagnosis of human ocular infection with T. gondii.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2017 |
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| Erschienen: |
2017 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:115 |
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| Enthalten in: |
Research in veterinary science - 115(2017) vom: 01. Dez., Seite 490-495 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Garcia, João Luis [VerfasserIn] |
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| Links: |
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| Themen: |
Animals |
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| Anmerkungen: |
Date Completed 29.05.2018 Date Revised 02.12.2018 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.rvsc.2017.07.022 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM274613662 |
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| 100 | 1 | |a Garcia, João Luis |e verfasserin |4 aut | |
| 245 | 1 | 4 | |a The use of ELISA, nPCR and qPCR for diagnosis of ocular toxoplasmosis in experimentally infected pigs |
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| 500 | |a Date Revised 02.12.2018 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2017 Elsevier Ltd. All rights reserved. | ||
| 520 | |a In the present study we experimentally infected pigs with T. gondii tachyzoites, bradyzoites and oocysts in order to evaluate IgG-ELISA, nested-PCR, and qPCR for diagnosis of ocular infection. Eighteen pigs were divided into four groups: G1 (infected with 103 tissue cysts of the M4 strain (type II) at day 28, n=5), G2 (infected with 103 oocysts of the M4 strain at day 28, n=5), G3 (infected with tachyzoites of S48 strain (type 1) at day 0, n=5), and G4 (uninfected unchallenged, control group n=3). At day 70 of the experiment all animals were culled, and serum, aqueous humor (AH) and vitreous humor (VH) samples were collected to perform indirect ELISA, and PCR (nPCR, and qPCR). By ELISA nine pigs (60%) out of 15 were positive in VH samples, and seven out of 15 (46%) were positive in AH samples. Both molecular techniques used here, nPCR and qPCR, were able to detect <50fg of T. gondii tachyzoite DNA. The nPCR and qPCR detected six (7/15, 47%) and two (2/15, 13.3%) positive animals respectively. Antibody responses were detected in serum and in AH and VH from the eye, suggesting that pigs may be an animal that could be used as a model to further our understanding of diagnosis of human ocular infection with T. gondii | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Animals | |
| 650 | 4 | |a Choroid | |
| 650 | 4 | |a Ocular toxoplasmosis | |
| 650 | 4 | |a Retina | |
| 650 | 4 | |a Toxoplasma gondii | |
| 650 | 7 | |a Antibodies, Protozoan |2 NLM | |
| 700 | 1 | |a Burrells, Alison |e verfasserin |4 aut | |
| 700 | 1 | |a Bartley, Paul M |e verfasserin |4 aut | |
| 700 | 1 | |a Bartley, Kathryn |e verfasserin |4 aut | |
| 700 | 1 | |a Innes, Elisabeth A |e verfasserin |4 aut | |
| 700 | 1 | |a Katzer, Frank |e verfasserin |4 aut | |
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