Xu Chao, Wei Qing-kuan, Li Jin, Xiao Ting, Yin Kun, Wang Yong-bin, Kong Xiang-li, Xu Yan, Cui Yong, Sun Hui, Zhu Song, Yan Ge, Huang Bing-cheng
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2016 Dec;34(6):482-8.
To investigate the mutation of genes associated with drug resistance (Pfcrt, Pfmdr1, Pfdhfr and K13） in imported Plasmodium falciparum in Shandong Province.
Blood was collected from 94 falciparum malaria cases who returned from Africa in 2014. Genomic DNA for P. falciparum was extracted from the blood samples and nested PCR was performed using primers specifically designed for Pfcrt, Pfmdr1, Pfdhfr and K13. The PCR products were sequenced. Gene mutations were analyzed by sequence alignment.
The 94 imported cases were from 18 African countries. Nested PCR was successful on DNA from all the blood samples except for Pfcrt amplification in one sample. Sequence analysis revealed three types of mutations Pfcrt K76T （36.6%, 34/93）, Pfmdr1 N86Y （21.3%, 20/94）, and Pfdhfr S108N （98.9%, 93/94） （χ2=127.5， P<0.05）. K13 C580Y mutation was not found. Co-occurrence of K76T, N86Y, and S108N was found in 6 blood samples （6.5%）, which were imported from Liberia（2）, Angola（1）, Equatorial guinea（1）, Congo（1）, and Guinea（1）. Co-occurrence of K76T and S108N mutations was found in 28 samples（30.1%）, and that of N86Y and S108N in 14 samples （15.1%）. Forty-four samples（47.3%） harbored S108N mutation only, and one sample was null for any of the mutations.
There are mutations in Pfcrt, Pfmdr1, and Pfdhfr in imported Plasmodium falciparum in Shandong Province. No mutation was found for the K13 gene.
调查山东省输入性恶性疟原虫中与耐药相关基因（Pfcrt、Pfmdr1、Pfdhfr和K13）的突变情况。
采集2014年从非洲回国的94例恶性疟疾病例的血液。从血样中提取恶性疟原虫的基因组DNA，并使用专门为Pfcrt、Pfmdr1、Pfdhfr和K13设计的引物进行巢式PCR。对PCR产物进行测序。通过序列比对分析基因突变情况。
94例输入性病例来自18个非洲国家。除1份样本的Pfcrt扩增外，所有血样的DNA巢式PCR均成功。序列分析显示三种突变类型：Pfcrt K76T（36.6%，34/93）、Pfmdr1 N86Y（21.3%，20/94）和Pfdhfr S108N（98.9%，93/94）（χ2=127.5，P<0.05）。未发现K13 C580Y突变。在6份血样（6.5%）中发现K76T、N86Y和S108N同时存在，这些血样分别来自利比里亚（2份）、安哥拉（1份）、赤道几内亚（1份）、刚果（1份）和几内亚（1份）。在28份样本（30.1%）中发现K76T和S108N突变同时存在，在14份样本（15.1%）中发现N86Y和S108N突变同时存在。44份样本（47.3%）仅存在S108N突变，1份样本未发现任何突变。
山东省输入性恶性疟原虫的Pfcrt、Pfmdr1和Pfdhfr存在突变。未发现K-13基因的突变。
