Two experiments were conducted to evaluate duodenal gene expression of lactoferrin and effect of weaning age on mRNA expression of lactoferrin in piglets using semi-quantitative RT-PCR analysis. In experiment 1, a total of 15 female Duroc x Landrace x Yorkshire piglets of five groups, each group pigs at 1, 14, 28, 42 and 56 days of age were used to determine developmental gene expression of lactoferrin in duodenum. In experiment 2, a total of 18 female neonatal piglets were divided into three groups, which were weaned at 21, 28 and 35 days of age respectively. In each group, three piglets' duodena were sampled at 21, 28 and 35 days of age and the other three piglets' duodena were sampled 7 days after weaning in each group. The samples were collected for detecting the effect of weaning age on lactoferrin mRNA expression of piglets. The results show that lactoferrin mRNA levels decreased steadily in postnatal day 1-56. But only from day 28-42 (14 days after weaned), the levels of lactoferrin gene expression were decreased markedly (p < 0.05), and the difference of lactoferrin mRNA levels at other stages was not significant. This result suggested that weaning had an effect on gene expression of lactoferrin. The results of experiment 2 showed that when the piglets were weaned at 21-28 and 35 days of age respectively, the expression levels of lactoferrin were decreased by 77%, 53% and 59% at the seventh day after weaning. Our results showed that weaning significantly decreased lactoferrin mRNA expression of piglets.