利用微衛星標幟進行臺灣純種及雜交（肉）豬產銷履歷追溯可能性之探討

Preliminary Study for Molecular Traceability of Pure and Hybrid Pigs (Fresh Pork) Using Microsatellite Markers in Taiwan

王佩華(P. H. Wang) ； 呂岳錚(Y. J. Lyu) ； 黃三元(S. Y. Huang) ； 簡執中(C. C. Chien) ； 杜清富(C. F. Tu) ； 林恩仲(E.-C. Lin)

《台灣農學會報》 13卷3期 (2012/06) Pp. 296-317

https://doi.org/10.6730/JAAT.201206_13(3).0005

摘要

近年來由於狂牛病及各種食品安全事件的爆發，使得食品安全日漸受到重視，歐盟會員國及世界各國都在推行產銷履歷制度以增進食品安全，而利用微衛星分子標幟進行遺傳追溯對傳統產銷履歷方法進行驗證成為近年來的研究方向之一，本研究之目的即在利用微衛星標幟建立臺灣地區（肉）豬隻產銷履歷追溯可能性之探討。本研究試驗動物來自中央檢定北站之三個品種的純種豬隻（41頭藍瑞斯、95頭約克夏及76頭杜洛克），共計212頭；以及來自27頭二品種雜交肉豬（藍瑞斯×杜洛克, LD），19頭三品種雜交肉豬（藍瑞斯×約克夏×杜洛克, LYD），研究採用10組微衛星標幟進行試驗。試驗結果顯示，三個純品種豬以10組微衛星標幟檢測，其理論異質度（HE）在無遺傳相關個體上，檢測結果分別為0.62、0.65及0.65；在所有個體間（部分具有遺傳相關）分別為0.61、0.66及0.65。其多態性訊息含量（PIC）在無遺傳相關個體上分別為0.57、0.60及0.60；在所有個體間（部分具有遺傳相關）分別為 0.57、0.6及0.61。10組微衛星標幟進行哈溫平衡（HWE）檢定的結果，在無遺傳相關個體上，藍瑞斯及約克夏各有一個標幟，杜洛克有三個標幟偏離HWE（P＜0.05），在所有個體間（部分具有遺傳相關），藍瑞斯、約克夏及杜洛克分別有一、二及三個標幟偏離 HWE（P＜0.05），但三個純品種豬無論個體有無遺傳相關，在10個微衛星標幟之tP（ID）並無顯著性差異。進行個體鑑定之結果，三個純品種豬其tP（ID）在遺傳相關個體上分別為1.42×10 -8、6.49×10 -9及1.23×10 -8；在所有個體間分別為1.58×10 -8、5.61×10 -9及9.34×10 -9，而對雜交豬隻進行個體鑑別之結果，LD、LYD雜交肉豬其tP（ID）分別為1.79×10 -9及6.14×10 -9，對偏離HWE之標幟之P（ID）進行修正後之tP（ID）sibs亦接近千萬分之一，因此不論純種或雜交肉豬，以此10個微衛星標幟所檢測的預估tP（ID）值均小於1×10 -7。綜上所述，臺灣近五年之每年上市屠宰的純種淘汰豬及雜交肉豬總數在一千萬頭以下，故屠宰豬在無論有無遺傳相關的情況下，此10組微衛星標幟應可達成臺灣常見之純種豬及雜交肉豬的個體鑑別需求，以應用於豬肉之產銷履歷追溯及檢測技術之建立。

關鍵字

豬微衛星標幟 ； 產銷履歷 ； 個體鑑別

並列摘要

In the last few decades, Bovine spongiform encephalopathy (BSE) and other food safety issues have aroused consumers' concern. EU member countries and other nations have regulations to ensure food traceability in order to promote food safety. Molecular traceability using DNA markers has become popular in recent years. Microsatellite markers have many advantages and are thus often utilized as molecular markers in molecular traceability. The objective of this study is to establish the microsatellite marker sets for genetic verification traceability of pure and hybrid pigs and fresh pork in Taiwan. In this study, purebred pigs of three breeds, total 212 heads (41 Landrace, 95 Yorkshire and 76 Duroc) from the central test station and 46 heads commercial crossbred pigs (27 Landrace×Duroc, LD, 19 Landrace×Yorkshire×Duroc, LYD), were genotyped using 10 microsatellite marker sets. We performed two multiplex polymerase chain reaction (PCR) with fluorescent-labeled microsatellite markers. The results showed that the expected heterozygosity (H E) in genetically unrelated individuals of three pure breeds were 0.62, 0.65 and 0.65, respectively; while that in all individuals (with partial genetic relationship) of three pure breeds were 0.61, 0.66 and 0.65, respectively. The polymorphism information content (PIC) in unrelated individuals of the three breeds were 0.57, 0.60 and 0.60, respectively; in all individuals (partial genetic relationship) of three breeds were 0.57, 0.61 and 0.61. After performing Hardy-Weinberg equilibrium (HWE) test in the samples, in unrelated individuals of the three breeds, there were one microsatellite marker in Landrace, one marker in Yorkshire and three markers in Duroc against HWE, respectively (P ＜ 0.05); in all individuals (partial genetic relationship) of three breeds, there were one marker in Landrace, two markers in Yorkshire and three in Duroc against HWE, respectively (P＜0.05). The results of individual identification showed that total probability of identify (tP (ID)) in unrelated individuals of three breeds were 1.42×10 -8 , 6.49×10 -9 and 1.23×10 -8 , respectively; in all individuals (partial genetic relationship) of three breeds were 1.58×10 -8 , 5.61×10 -9 and 9.34×10 -9 , respectively. The tP (ID) of LD and LYD crossbred pigs were 1.79×10 -9 and 6.14×10 -9, respectively. All the tested groups had their tP (ID) or probability of identify in sibs (tP (ID) sibs) values less than 1.00×10 -7. In Taiwan, the annual number of purebred and crossbred pigs slaughtered was less than 10 million (1×10 7) heads in the past five years. Thus, according to the results of this study, 10 microsatellite markers are sufficient for individual identification among the common purebred and crossbred commercial pigs in the molecular traceability of purebred and crossbred pigs as well as their fresh pork.