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Quantification of hookworm ova from wastewater matrices using quantitative PCR

Pradip Gyawali  Warish Ahmed  Jatinder P.Sidhu  Paul Jagals  Simon Toze  
【摘要】:A quantitative PCR(q PCR) assay was used to quantify Ancylostoma caninum ova in wastewater and sludge samples.We estimated the average gene copy numbers for a single ovum using a mixed population of ova.The average gene copy numbers derived from the mixed population were used to estimate numbers of hookworm ova in A.caninum seeded and unseeded wastewater and sludge samples.The newly developed qP CR assay estimated an average of3.7 × 10~3 gene copies per ovum,which was then validated by seeding known numbers of hookworm ova into treated wastewater.The qP CR estimated an average of(1.1 ± 0.1),(8.6 ± 2.9)and(67.3 ± 10.4) ova for treated wastewater that was seeded with(1 ± 0),(10 ± 2) and(100 ± 21)ova,respectively.The further application of the q PCR assay for the quantification of A.caninum ova was determined by seeding a known numbers of ova into the wastewater matrices.The qP CR results indicated that 50%,90% and 67% of treated wastewater(1 L),raw wastewater(1 L)and sludge(~4 g) samples had variable numbers of A.caninum gene copies.After conversion of the q PCR estimated gene copy numbers to ova for treated wastewater,raw wastewater,and sludge samples,had an average of 0.02,1.24 and 67 ova,respectively.The result of this study indicated that qP CR can be used for the quantification of hookworm ova from wastewater and sludge samples;however,caution is advised in interpreting qP CR generated data for health risk assessment.

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