Amplification of T. gondii specific nucleotide sequences were attempted on gel purified (see methods) DNA extracts from 25 portions of beef, chicken, lamb and pork mince purchased from butchers and supermarkets from within the Hamilton area. The results are summarised in Tables 3.3 to 3.26 below.
Genotyping of T. gondii from commercially available meat samples using amplified polymorphic PCR targets such as SAG3, L368 and GRA6 was not possible, either owing to non-specific primer binding, the complete lack of T. gondii template DNA or extremely low T. gondii template concentration. Overall these results show that if T. gondii is present in NZ farmed meat for sale in the Hamilton region, cyst density is lower than would have been predicted by overseas studies conducted in the United Kingdom (Aspinall et al., 2002) and in the USA (J. Dubey et al., 2005).
Table 3.8 A summary of Beef1 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions. (seq+) and (seq-) indicate positive and negative sequencing results (see Appendix 2 for detail).
Table 3.9 A summary of Lamb1 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions. (seq+) and (seq-) indicate positive and negative sequencing results (see Appendix 2 for detail).
Table 3.10 A summary of Pork1 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions. (seq+) and (seq-) indicate positive and negative sequencing results (see appendix for detail).
Although the original Pork1 extract tested positive for T. gondii DNA, a further 3 extracts were carried out (approximately 0.5 grams per extraction), no T. gondii DNA was detected using both nested and conventional PCR reactions.
Table 3.11 A summary of Beef2 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. (seq+) or (seq-) show sequencing results. Bracketed numbers show nested PCR reactions.
Table 3.12 A summary of Lamb2 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions. (seq+) and (seq-) indicate positive and negative sequencing results.
Table 3.13 A summary of Pork2 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions. (seq+) and (seq-) indicate positive and negative sequencing results.
Table 3.14 A summary of Beef3 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions.
Table 3.15 A summary of Chicken3 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions. (seq+) and (seq-) show positive and negative sequencing results (see Appendix 2 for detail).
Table 3.16 A summary of Lamb3 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. (seq+) or (seq-) show sequencing results (see Appendix 2 for detail). Bracketed numbers indicate nested PCR reactions.
Table 3.17 A summary of Pork3 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions.
Table 3.18 A summary of Beef4 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions.
.
Table 3.19 A summary of Lamb4 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions.
Table 3.20 A summary of Pork4 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers indicate nested PCR reactions.
Table 3.21 A summary of Beef5 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions. (seq+) and (seq-) show positive and negative sequencing results (see Appendix 2 for detailed sequence data).
Table 3.22 A summary of Chicken5 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions. (seq+) and (seq-) show positive and negative sequencing results
Table 3.23 A summary of Lamb5 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions. (seq+) and (seq-) show positive and negative sequencing results (see Appendix 2 for detailed sequence data).
Table 3.24 A summary of Pork5 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions.
Table 3.25 A summary of Beef6 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions.
Table 3.26 A summary of Lamb6 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions.
Table 3.27 A summary of Lamb6 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions.
Table 3.28 A summary of Beef7 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions.
Table 3.29 A summary of Chicken7 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions.
Table 3.30 A summary of Lamb7 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions
Table 3.31 A summary of Pork7 PCR results with each primer set and thermocycler protocol trialled. N= no visible product. MP= multiple product
bands visible. S= single product band visible. Bracketed numbers show nested PCR reactions.