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PHD : Parasites, Hosts and Diseases

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Original Article

Tissue Fluid Enzyme-Linked Immunosorbant Assay for Piglets Experimentally Infected with Toxoplasma gondii and Survey on Local and Imported Pork in Korean Retail Meat Markets

The Korean Journal of Parasitology 2018;56(5):437-446.
Published online: October 31, 2018

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14Department of Preventive Veterinary Medicine, College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang 163319, P. R. 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*Corresponding authors (hongsj@cau.ac.kr; younhj@snu.ac.kr)

These authors contributed equally to this work.

• Received: September 27, 2018   • Revised: October 18, 2018   • Accepted: October 19, 2018

Copyright © 2018 by The Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Tissue Fluid Enzyme-Linked Immunosorbant Assay for Piglets Experimentally Infected with Toxoplasma gondii and Survey on Local and Imported Pork in Korean Retail Meat Markets
Korean J Parasitol. 2018;56(5):437-446.   Published online October 31, 2018
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Tissue Fluid Enzyme-Linked Immunosorbant Assay for Piglets Experimentally Infected with Toxoplasma gondii and Survey on Local and Imported Pork in Korean Retail Meat Markets
Korean J Parasitol. 2018;56(5):437-446.   Published online October 31, 2018
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Tissue Fluid Enzyme-Linked Immunosorbant Assay for Piglets Experimentally Infected with Toxoplasma gondii and Survey on Local and Imported Pork in Korean Retail Meat Markets
Image Image Image
Fig. 1 Increase of anti-T. gondii antibodies in the sera of standard positive controls in the early phase of infection.
Fig. 2 Antibody titer of each part of the T. gondii-infected piglets using CAU-tf-ELISA kit and Commercial kit S. (A) The mean±2 standard deviations of the antibody titer of Group D was designated as the cut-off value (0.15) for CAU-tf-ELISA. (B) The cut-off value (0.3) for the OD450 was used for the Commercial kit S according to the manufacturer’s guideline. Tissue-specific cut-off values were applied for CAU-tf-ELISA (C) and Commercial kit S (D) according to each meat portion (See the details in Table 3).
Fig. 3 Correlation between T. gondii antibody levels of the standard positive controls assayed using the CAU-tf-ELISA kit and Commercial kit S.
Tissue Fluid Enzyme-Linked Immunosorbant Assay for Piglets Experimentally Infected with Toxoplasma gondii and Survey on Local and Imported Pork in Korean Retail Meat Markets

Comparison of the functionality of CAU-tf-ELISA kit and Commercial kit S for tissue fluids samples from the standard positive controls

Pork Part No. of positive samples (%)a

Group A Group B Group C Subtotal (Groups A-C) Group D





CAU-tf-ELISA Commercial kit S CAU-tf-ELISA Commercial kit S CAU-tf-ELISA Commercial kit S CAU-tf-ELISA Commercial kit S CAU-tf-ELISA Commercial kit S
Heart 2 (100) 2 (100) 4 (100) 4 (100) 4 (100) 1 (25.0) 10 (100) 7 (70.0) 1 (33.3) 0 (0.0)

Skirt meat 2 (100) 2 (100) 4 (100) 4 (100) 3 (75.0) 1 (25.0) 9 (90.0) 7 (70.0) 0 (0.0) 0 (0.0)

Tenderloin 2 (100) 2 (100) 4 (100) 3 (75.0) 3 (75.0) 0 (0.0) 9 (90.0) 5 (50.0) 0 (0.0) 0 (0.0)

Sirloin 2 (100) 2 (100) 4 (100) 0 (0.0) 1 (25.0) 0 (0.0) 7 (70.0) 2 (20.0) 0 (0.0) 0 (0.0)

Pork belly 2 (100) 1 (50.0) 4 (100) 0 (0.0) 1 (25.0) 0 (0.0) 7 (70.0) 1 (10.0) 0 (0.0) 0 (0.0)

Shoulder picnic 2 (100) 1 (50.0) 4 (100) 0 (0.0) 0 (0.0) 0 (0.0) 6 (60.0) 1 (10.0) 0 (0.0) 0 (0.0)

Ham 2 (100) 2 (100) 3 (75.0) 1 (25.0) 0 (0.0) 0 (0.0) 5 (50.0) 3 (30.0) 0 (0.0) 0 (0.0)

Shoulder blade 2 (100) 1 (50.0) 3 (75.0) 0 (0.0) 1 (25.0) 0 (0.0) 6 (60.0) 1 (10.0) 0 (0.0) 0 (0.0)

Total 16 (100) 13 (81.3) 30 (93.8) 12 (37.5) 13 (40.6) 2 (6.3) 59 (73.8) 27 (33.8) 1 (3.3) 0 (0.0)

Serumb 2 (100) 2 (100) 4 (100) 4 (100) 4 (100) 1 (25.0) 10 (100) 7 (70.0) 1 (3.3) 0 (0.0)

aPositive rate (%) was determined by a cut-off value of 0.15 for the CAU-tf-ELISA kit and 0.3 for the Commercial kit S.

bAs supporting data.

Diagnostic performance of the CAU-tf-ELISA kit and Commercial kit S in terms of detection of anti-T. gondii antibodies

ELISA (cut-off value) Results No. of specimens with Diagnostic performance [95% confidence interval]


Positive Negative Subtotal Sensitivity Specificity Positive predictive value Negative predictive value
CAU-tf-ELISA (0.15) Positive 69 2 71
Negative 21 25 46
Subtotal 90 27 117 76.70% [66.6% to 84.9%] 92.60% [75.7% to 99.1%] 97.20% [90.1% to 99.3%] 54.40% [44.6% to 63.7%]

Commercial kit S (0.3) Positive 34 0 34
Negative 56 27 83
Subtotal 90 27 117 37.80% [27.8% to 48.6%] 100% [87.2% to 100%] 100% 32.50% [29.1% to 36.2%]

CAU-tf-ELISA (tissue-specific values) Positive 87 0 87
Negative 3 27 30
Subtotal 90 27 117 96.70% [90.6% to 99.3%] 100% [87.2% to 100%] 100% 90.00% [74.7% to 96.5%]

Functionality of CAU-tf-ELISA kit and Commercial kit S applied with tissue-specific cut-off values

Pork Part CAU-tf-ELISA Commercial kit S


Cut-off value No. of positive (%) Cut-off value No. of positive (%)


Groups A-C Group D Groups A-C Group D
Heart 0.199 10 (100) 0 (0.0) 0.005 10 (100) 2 (66.6)

Skirt meat 0.068 10 (100) 0 (0.0) 0.072 9 (90.0) 0 (0.0)

Tenderloin 0.088 10 (100) 0 (0.0) 0.006 10 (100) 0 (0.0)

Sirloin 0.073 10 (100) 0 (0.0) 0.005 9 (90.0) 0 (0.0)

Belly 0.079 10 (100) 0 (0.0) 0.006 10 (100) 0 (0.0)

Shoulder picnic 0.074 10 (100) 0 (0.0) 0.005 9 (90.0) 1 (33.3)

Ham 0.115 7 (70.0) 0 (0.0) 0.016 7 (70.0) 0 (0.0)

Shoulder blade 0.066 10 (100) 0 (0.0) 0.006 9 (90.0) 0 (0.0)

Total 77 (96.3) 0 (0.0) 73 (91.3) 43 (12.5)

Seruma 0.207 10 (100) 0 (0.0) 0.005 10 (100) 1 (33.3)

aAs supporting data.

Prevalence of anti-T. gondii antibodies in the tissue fluid of local and imported pork determined by CAU-tf-ELISA

Pork No. of samples collected Positivity (%)

Meat Tissue fluid
Local
 Skirt meat 120 116 44 (37.9)
 Heart 120 120 8 (6.7)
 Belly 124 117 1 (0.9)
 Shoulder picnic 120 113 0 (0.0)
 Loin 120 117 0 (0.0)
Subtotal 604 583 53 (9.1)

Imported (Country)
Bellya 186 183 0 (0.0)
 Skirt meat (Mexico) 20 20 2 (10.0)
 Skirt meat (Canada) 20 20 0 (0.0)
 Skirt meat (France) 20 20 0 (0.0)
 Shoulder picnic (USA) 22 22 0 (0.0)
 Shoulder blade (Chile) 62 62 0 (0.0)
 Shoulder blade (Mexico) 50 49 4 (8.2)
 Shoulder blade (Spain) 10 10 1 (10.0)
Subtotal 390 386 7 (1.8)

Total 994 969 60 (6.2)

aTwenty imported pork belly samples analyzed from Austria, Belgium, USA, Chile, France, Hungary, the Netherlands, and Spain each, and 26 samples from Germany were analyzed.

Prevalence of T. gondii antibodies in the tissue fluid of local pork using CAU-tf-ELISA according to the surveyed region

Pork No. examined No. of positive samples (%)
Seoul Daejeon Daegu Busan Gwangju Jeju Subtotal
Skirt 116 2 (10.0) 3 (15.8) 10 (58.8) 12 (60.0) 6 (30.0) 11 (55.0) 44 (37.9)
Heart 120 0 (0.0) 0 (0.0) 3 (15.0) 3 (15.0) 1 (5.0) 1 (5.0) 8 (6.7)
Belly 117 0 (0.0) 1 (5.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 1 (0.9)
Shoulder picnic 113 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0)
Loin 117 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0)
Total 583 2 (2.0) 4 (4.0) 13 (15.1) 15 (15.5) 7 (7.0) 12 (11.9) 53 (9.1)
Table 1 Comparison of the functionality of CAU-tf-ELISA kit and Commercial kit S for tissue fluids samples from the standard positive controls

Positive rate (%) was determined by a cut-off value of 0.15 for the CAU-tf-ELISA kit and 0.3 for the Commercial kit S.

As supporting data.

Table 2 Diagnostic performance of the CAU-tf-ELISA kit and Commercial kit S in terms of detection of anti-T. gondii antibodies
Table 3 Functionality of CAU-tf-ELISA kit and Commercial kit S applied with tissue-specific cut-off values

As supporting data.

Table 4 Prevalence of anti-T. gondii antibodies in the tissue fluid of local and imported pork determined by CAU-tf-ELISA

Twenty imported pork belly samples analyzed from Austria, Belgium, USA, Chile, France, Hungary, the Netherlands, and Spain each, and 26 samples from Germany were analyzed.

Table 5 Prevalence of T. gondii antibodies in the tissue fluid of local pork using CAU-tf-ELISA according to the surveyed region