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Molecular mutation profile of pfcrt in Plasmodium falciparum isolates imported from Africa in Henan province.

Zhou RM, Zhang HW, Yang CY, Liu Y, Zhao YL, Li SH, Qian D, Xu BL - Malar. J. (2016)

Bottom Line: Four haplotypes coding 72-76 of pfcrt were found including CVMNK (wild type), CVIET (mutation type), CVIEK (mutation type), and CV M/I N/E/D/K K/T (mixed type), with 61.95 % (311/502), 33.07 % (166/502), 0.20 % (1/502), and 4.78 % (24/502) prevalence, respectively.There was significant difference among the groups (χ(2) = 23.78, P < 0.05).There was no significant difference among the groups (χ(2) = 2.31, P > 0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Parasite Disease Control and Prevention, Henan Province Center for Disease Control and Prevention, Zhengzhou, 450016, People's Republic of China.

ABSTRACT

Background: Anti-malarial drug resistance is a primary public health problem. Haplotypes of pfcrt gene have been implicated to be molecular markers of chloroquine (CQ) resistance. This study aims to explore the prevalence of polymorphisms in pfcrt in Plasmodium falciparum-infected patients imported from Africa in Henan province.

Methods: Blood samples were collected from 502 patients who were infected with P. falciparum returning from Africa in Henan province during 2012-2015. The single nucleotide polymorphisms in pfcrt (codons 72-76) were assessed by nested PCR with DNA sequencing and restriction digestion, the haplotype prevalences were also determined.

Results: Four haplotypes coding 72-76 of pfcrt were found including CVMNK (wild type), CVIET (mutation type), CVIEK (mutation type), and CV M/I N/E/D/K K/T (mixed type), with 61.95 % (311/502), 33.07 % (166/502), 0.20 % (1/502), and 4.78 % (24/502) prevalence, respectively. Except mixed type, CVIET and CVIEK were the largest proportion of the mutant type in West Africa, accounting for 44.83 % (91/203), followed by East Africa (8/21, 38.10 %), North Africa (4/11, 36.36 %), Central Africa (36/135, 26.67 %), and South Africa (28/132, 21.21 %). There was significant difference among the groups (χ(2) = 23.78, P < 0.05). Mixed type was the largest proportion in North Africa (9.09 %), followed by Central Africa (6.67 %), East Africa (4.76 %), South Africa (4.55 %), and West Africa (3.45 %). There was no significant difference among the groups (χ(2) = 2.31, P > 0.05). The position 72 and 73 of pfcrt showed predominance for the wild type with rates of 100 % (502/502).

Conclusions: This study identified four haplotypes of pfcrt in P. falciparum-infected patients imported from Africa in Henan province. The prevalence of mutations in the pfcrt was dropped comparing with other people's researches. It establishes fundamental data for detection of P. falciparum CQR with molecular markers for the imported P. falciparum in China, and it also provides complementary information of CQR for the malaria endemic countries and assesses the evolution of anti-malarial drug resistance.

No MeSH data available.


Related in: MedlinePlus

Identification of the four haplotypes by enzyme digestion. Lanes1, 3, 5 and 7 were the nested PCR product of the four haplotypes. Lanes2, 4, 6 and 8 were digested by Apo I. M, molecular marker; lanes1 and 2, CVMNK; lanes3 and 4, CVM/I N/E/D/K T/K; lanes5 and 6, CVIET; lanes7 and 8, CVIEK
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Fig1: Identification of the four haplotypes by enzyme digestion. Lanes1, 3, 5 and 7 were the nested PCR product of the four haplotypes. Lanes2, 4, 6 and 8 were digested by Apo I. M, molecular marker; lanes1 and 2, CVMNK; lanes3 and 4, CVM/I N/E/D/K T/K; lanes5 and 6, CVIET; lanes7 and 8, CVIEK

Mentions: The nested PCR yielded a 145-bp PCR product for each of the 502 samples. These products contained the codons of interest, 72 through 76, which were analysed both by enzymatic digestion and sequencing. The resulting of Apo I enzymatic digestion yielded two fragments of 31 and 114 bp in the case of the parasite strains containing the wild K76 variant, whereas the mutant T76 variant yielded only one fragment since there was no digestion. Multi-clonal variants yielded three fragments (145, 114 and 31-bp products). A photo of agarose gel electrophoresis is shown in Fig. 1. Restriction digest of the 502 samples gave the following results: 166 (33.07 %) mutant types, 312 (62.15 %) wild types and 24 (4.78 %) mixed infections, which were also confirmed by sequencing (see Fig. 2).Fig. 1


Molecular mutation profile of pfcrt in Plasmodium falciparum isolates imported from Africa in Henan province.

Zhou RM, Zhang HW, Yang CY, Liu Y, Zhao YL, Li SH, Qian D, Xu BL - Malar. J. (2016)

Identification of the four haplotypes by enzyme digestion. Lanes1, 3, 5 and 7 were the nested PCR product of the four haplotypes. Lanes2, 4, 6 and 8 were digested by Apo I. M, molecular marker; lanes1 and 2, CVMNK; lanes3 and 4, CVM/I N/E/D/K T/K; lanes5 and 6, CVIET; lanes7 and 8, CVIEK
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4862149&req=5

Fig1: Identification of the four haplotypes by enzyme digestion. Lanes1, 3, 5 and 7 were the nested PCR product of the four haplotypes. Lanes2, 4, 6 and 8 were digested by Apo I. M, molecular marker; lanes1 and 2, CVMNK; lanes3 and 4, CVM/I N/E/D/K T/K; lanes5 and 6, CVIET; lanes7 and 8, CVIEK
Mentions: The nested PCR yielded a 145-bp PCR product for each of the 502 samples. These products contained the codons of interest, 72 through 76, which were analysed both by enzymatic digestion and sequencing. The resulting of Apo I enzymatic digestion yielded two fragments of 31 and 114 bp in the case of the parasite strains containing the wild K76 variant, whereas the mutant T76 variant yielded only one fragment since there was no digestion. Multi-clonal variants yielded three fragments (145, 114 and 31-bp products). A photo of agarose gel electrophoresis is shown in Fig. 1. Restriction digest of the 502 samples gave the following results: 166 (33.07 %) mutant types, 312 (62.15 %) wild types and 24 (4.78 %) mixed infections, which were also confirmed by sequencing (see Fig. 2).Fig. 1

Bottom Line: Four haplotypes coding 72-76 of pfcrt were found including CVMNK (wild type), CVIET (mutation type), CVIEK (mutation type), and CV M/I N/E/D/K K/T (mixed type), with 61.95 % (311/502), 33.07 % (166/502), 0.20 % (1/502), and 4.78 % (24/502) prevalence, respectively.There was significant difference among the groups (χ(2) = 23.78, P < 0.05).There was no significant difference among the groups (χ(2) = 2.31, P > 0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Parasite Disease Control and Prevention, Henan Province Center for Disease Control and Prevention, Zhengzhou, 450016, People's Republic of China.

ABSTRACT

Background: Anti-malarial drug resistance is a primary public health problem. Haplotypes of pfcrt gene have been implicated to be molecular markers of chloroquine (CQ) resistance. This study aims to explore the prevalence of polymorphisms in pfcrt in Plasmodium falciparum-infected patients imported from Africa in Henan province.

Methods: Blood samples were collected from 502 patients who were infected with P. falciparum returning from Africa in Henan province during 2012-2015. The single nucleotide polymorphisms in pfcrt (codons 72-76) were assessed by nested PCR with DNA sequencing and restriction digestion, the haplotype prevalences were also determined.

Results: Four haplotypes coding 72-76 of pfcrt were found including CVMNK (wild type), CVIET (mutation type), CVIEK (mutation type), and CV M/I N/E/D/K K/T (mixed type), with 61.95 % (311/502), 33.07 % (166/502), 0.20 % (1/502), and 4.78 % (24/502) prevalence, respectively. Except mixed type, CVIET and CVIEK were the largest proportion of the mutant type in West Africa, accounting for 44.83 % (91/203), followed by East Africa (8/21, 38.10 %), North Africa (4/11, 36.36 %), Central Africa (36/135, 26.67 %), and South Africa (28/132, 21.21 %). There was significant difference among the groups (χ(2) = 23.78, P < 0.05). Mixed type was the largest proportion in North Africa (9.09 %), followed by Central Africa (6.67 %), East Africa (4.76 %), South Africa (4.55 %), and West Africa (3.45 %). There was no significant difference among the groups (χ(2) = 2.31, P > 0.05). The position 72 and 73 of pfcrt showed predominance for the wild type with rates of 100 % (502/502).

Conclusions: This study identified four haplotypes of pfcrt in P. falciparum-infected patients imported from Africa in Henan province. The prevalence of mutations in the pfcrt was dropped comparing with other people's researches. It establishes fundamental data for detection of P. falciparum CQR with molecular markers for the imported P. falciparum in China, and it also provides complementary information of CQR for the malaria endemic countries and assesses the evolution of anti-malarial drug resistance.

No MeSH data available.


Related in: MedlinePlus