Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in pork
Trichinella is a zoonotic nematode parasite transmitted by the ingestion of raw or under-cooked meat. Control of the parasite is essential to facilitate public health and trade in products from susceptible food animals, including pork and horse meat. The standard method for detecting Trichinella mus...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-03-01
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| Series: | Food and Waterborne Parasitology |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405676617300197 |
| _version_ | 1819162295306027008 |
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| author | Alvin Gajadhar Kelly Konecsni Brad Scandrett Patrik Buholzer |
| author_facet | Alvin Gajadhar Kelly Konecsni Brad Scandrett Patrik Buholzer |
| author_sort | Alvin Gajadhar |
| collection | DOAJ |
| description | Trichinella is a zoonotic nematode parasite transmitted by the ingestion of raw or under-cooked meat. Control of the parasite is essential to facilitate public health and trade in products from susceptible food animals, including pork and horse meat. The standard method for detecting Trichinella muscle larvae uses pepsin enzyme and hydrochloric acid (HCl) in an artificial digestion procedure. A new artificial digestion assay using serine protease was recently developed and commercialized (PrioCHECK™ Trichinella AAD) for the detection of Trichinella larvae in the muscle of infected animals. The assay uses no hazardous substances such as HCl or pepsin. Activation of the enzyme requires an elevated digestion temperature of 60 °C which kills the parasite and reduces the risk of contaminating the environment with Trichinella. Compared to the pepsin/HCl method, digestion time for the PrioCHECK Trichinella AAD assay is reduced by a third. A recent study demonstrated these features of the new assay and its suitability for digesting various muscles from domestic and wild animals. To further validate the assay's performance relative to the conventional pepsin/HCl digestion method several comparative studies were conducted using samples from different muscle sites spiked with low levels of encapsulated first stage Trichinella larvae (L1). Multiple muscle samples were collected from diaphragm, tongue, masseter, and loin of 3–4 month old pigs. Samples were spiked with 3, 4, 5, or 25 Trichinella spiralis L1. A total of 320 meat samples of 100 g each were used to compare the diagnostic proficiency of the PrioCHECK Trichinella AAD assay with the pepsin/HCl digestion method. Comparative and validation data produced from these studies showed that both methods are capable of consistently detecting Trichinella in 100 g samples which contained as few as 3 L1 or 0.03 larvae per gram of meat. Overall, the PrioCHECK Trichinella AAD assay performed satisfactorily according to international guidelines of the World Organization for Animal Health (OIE), European Union (EU) and International Commission on Trichinellosis (ICT) for the detection of Trichinella infection in pork. Keywords: Pork, Food safety, PrioCHECK Trichinella AAD assay, Pepsin/HCl digestion method, Diagnostic assay, Validation |
| first_indexed | 2024-12-22T17:25:58Z |
| format | Article |
| id | doaj.art-19567f7b6174411eb3bab428f1fd487b |
| institution | Directory Open Access Journal |
| issn | 2405-6766 |
| language | English |
| last_indexed | 2024-12-22T17:25:58Z |
| publishDate | 2018-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Food and Waterborne Parasitology |
| spelling | doaj.art-19567f7b6174411eb3bab428f1fd487b2022-12-21T18:18:42ZengElsevierFood and Waterborne Parasitology2405-67662018-03-0110613Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in porkAlvin Gajadhar0Kelly Konecsni1Brad Scandrett2Patrik Buholzer3Parasitix Lab Services, Innovation Place, Saskatoon, Sask. S7N 3R2, Canada; Corresponding author.Centre for Foodborne and Animal Parasitology, Canadian Food Inspection Agency, 116 Veterinary Road, Saskatoon, Sask. S7N 2R3, CanadaCentre for Foodborne and Animal Parasitology, Canadian Food Inspection Agency, 116 Veterinary Road, Saskatoon, Sask. S7N 2R3, CanadaThermo Fisher Scientific, Wagistrasse 27A, 8952 Schlieren, SwitzerlandTrichinella is a zoonotic nematode parasite transmitted by the ingestion of raw or under-cooked meat. Control of the parasite is essential to facilitate public health and trade in products from susceptible food animals, including pork and horse meat. The standard method for detecting Trichinella muscle larvae uses pepsin enzyme and hydrochloric acid (HCl) in an artificial digestion procedure. A new artificial digestion assay using serine protease was recently developed and commercialized (PrioCHECK™ Trichinella AAD) for the detection of Trichinella larvae in the muscle of infected animals. The assay uses no hazardous substances such as HCl or pepsin. Activation of the enzyme requires an elevated digestion temperature of 60 °C which kills the parasite and reduces the risk of contaminating the environment with Trichinella. Compared to the pepsin/HCl method, digestion time for the PrioCHECK Trichinella AAD assay is reduced by a third. A recent study demonstrated these features of the new assay and its suitability for digesting various muscles from domestic and wild animals. To further validate the assay's performance relative to the conventional pepsin/HCl digestion method several comparative studies were conducted using samples from different muscle sites spiked with low levels of encapsulated first stage Trichinella larvae (L1). Multiple muscle samples were collected from diaphragm, tongue, masseter, and loin of 3–4 month old pigs. Samples were spiked with 3, 4, 5, or 25 Trichinella spiralis L1. A total of 320 meat samples of 100 g each were used to compare the diagnostic proficiency of the PrioCHECK Trichinella AAD assay with the pepsin/HCl digestion method. Comparative and validation data produced from these studies showed that both methods are capable of consistently detecting Trichinella in 100 g samples which contained as few as 3 L1 or 0.03 larvae per gram of meat. Overall, the PrioCHECK Trichinella AAD assay performed satisfactorily according to international guidelines of the World Organization for Animal Health (OIE), European Union (EU) and International Commission on Trichinellosis (ICT) for the detection of Trichinella infection in pork. Keywords: Pork, Food safety, PrioCHECK Trichinella AAD assay, Pepsin/HCl digestion method, Diagnostic assay, Validationhttp://www.sciencedirect.com/science/article/pii/S2405676617300197 |
| spellingShingle | Alvin Gajadhar Kelly Konecsni Brad Scandrett Patrik Buholzer Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in pork Food and Waterborne Parasitology |
| title | Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in pork |
| title_full | Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in pork |
| title_fullStr | Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in pork |
| title_full_unstemmed | Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in pork |
| title_short | Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in pork |
| title_sort | validation of a new commercial serine protease artificial digestion assay for the detection of trichinella larvae in pork |
| url | http://www.sciencedirect.com/science/article/pii/S2405676617300197 |
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