Chloroquine resistant Plasmodium vivax

in vitro characterisation and association with molecular polymorphisms

Rossarin Suwanarusk, Bruce Russell, M Chavchich, Ferryanto Chalfein, Enny Kenangalem, V Kosaisavee, Prasetyorini, Kim Piera, Marion Barends, A Brockman, U Lek-uthai, Nicholas Anstey, Emiliana Tjitra, François Nosten, Q Cheng, Ric Price

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    Abstract

    Background: Treatment failure of chloroquine for P. vivax infections has reached high levels in the eastern provinces of Indonesia, however, in vitro characterization of chloroquine resistance and its associated molecular profile have yet to be determined.

    Methods: Using a modified schizont maturation assay we investigated the in vitro chloroquine susceptibility profile and molecular polymorphisms of P. vivax isolates collected from Papua, Indonesia, where high levels of clinical chloroquine treatment failure have been reported, and from Thailand, where chloroquine treatment is generally effective.

    Results: The geometric mean chloroquine IC50 for P. vivax isolates from Papua (n=145) was 312 nM [95%Cl: 237-411 nM] compared to 46.8 nM [95%Cl: 34.7-63.1 nM] from Thailand (n = 81); p<0.001. Correlating with the known clinical efficacy of the area, a cut off for chloroquine resistance was defined as 220nM, a level exceeded in 13.6% (11/81) of Thai isolates and 65% (94/145) of Papuan isolates; p<0.001. Several sequence polymorphisms in pvcrt-o and pvmdr1, and difference in pvmdr1 copy number were identified. A Y976F mutation in pvmdr1 was present in 96% (123/128) of Papuan isolates and 25% (17/69) of Thai isolates; p<0.001. Overall, the geometric mean chloroquine IC50 in isolates with the Y976F mutation was 283 nM [95%Cl: 211-379], compared to 44.5 nM [95%Cl: 31.3-63.4] in isolates with the wild type; p< 0.001. Pvmdr1 amplification occurred in 23% (15/66) of Thai isolates compared to none (0/104) of Indonesian isolates (p<0.001), but was not associated with increased chloroquine resistance after controlling for geographical location.

    Conclusions: In vitro susceptibility testing of P. vivax discriminates between populations with differing levels of clinical efficacy of chloroquine. The pvmdr1 polymorphism at Y976F may provide a useful tool to highlight areas of emerging chloroquine resistance, although further studies defining its clinical correlates are needed. � 2007 Suwanarusk et al.
    Original languageEnglish
    Article numbere1089
    Pages (from-to)1-9
    Number of pages9
    JournalPLoS One
    Volume2
    Issue number10
    DOIs
    Publication statusPublished - 31 Oct 2007

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    Plasmodium vivax
    chloroquine
    Chloroquine
    Polymorphism
    polymorphism
    Association reactions
    Indonesia
    Thailand
    Treatment Failure
    Inhibitory Concentration 50
    inhibitory concentration 50
    In Vitro Techniques
    Schizonts
    mutation
    schizonts
    Mutation
    Geographical Locations
    Malaria
    Amplification
    Assays

    Cite this

    Suwanarusk, R., Russell, B., Chavchich, M., Chalfein, F., Kenangalem, E., Kosaisavee, V., ... Price, R. (2007). Chloroquine resistant Plasmodium vivax: in vitro characterisation and association with molecular polymorphisms. PLoS One, 2(10), 1-9. [e1089]. https://doi.org/10.1371/journal.pone.0001089
    Suwanarusk, Rossarin ; Russell, Bruce ; Chavchich, M ; Chalfein, Ferryanto ; Kenangalem, Enny ; Kosaisavee, V ; Prasetyorini ; Piera, Kim ; Barends, Marion ; Brockman, A ; Lek-uthai, U ; Anstey, Nicholas ; Tjitra, Emiliana ; Nosten, François ; Cheng, Q ; Price, Ric. / Chloroquine resistant Plasmodium vivax : in vitro characterisation and association with molecular polymorphisms. In: PLoS One. 2007 ; Vol. 2, No. 10. pp. 1-9.
    @article{ef94e20f89e744359ec333478005a991,
    title = "Chloroquine resistant Plasmodium vivax: in vitro characterisation and association with molecular polymorphisms",
    abstract = "Background: Treatment failure of chloroquine for P. vivax infections has reached high levels in the eastern provinces of Indonesia, however, in vitro characterization of chloroquine resistance and its associated molecular profile have yet to be determined. Methods: Using a modified schizont maturation assay we investigated the in vitro chloroquine susceptibility profile and molecular polymorphisms of P. vivax isolates collected from Papua, Indonesia, where high levels of clinical chloroquine treatment failure have been reported, and from Thailand, where chloroquine treatment is generally effective. Results: The geometric mean chloroquine IC50 for P. vivax isolates from Papua (n=145) was 312 nM [95{\%}Cl: 237-411 nM] compared to 46.8 nM [95{\%}Cl: 34.7-63.1 nM] from Thailand (n = 81); p<0.001. Correlating with the known clinical efficacy of the area, a cut off for chloroquine resistance was defined as 220nM, a level exceeded in 13.6{\%} (11/81) of Thai isolates and 65{\%} (94/145) of Papuan isolates; p<0.001. Several sequence polymorphisms in pvcrt-o and pvmdr1, and difference in pvmdr1 copy number were identified. A Y976F mutation in pvmdr1 was present in 96{\%} (123/128) of Papuan isolates and 25{\%} (17/69) of Thai isolates; p<0.001. Overall, the geometric mean chloroquine IC50 in isolates with the Y976F mutation was 283 nM [95{\%}Cl: 211-379], compared to 44.5 nM [95{\%}Cl: 31.3-63.4] in isolates with the wild type; p< 0.001. Pvmdr1 amplification occurred in 23{\%} (15/66) of Thai isolates compared to none (0/104) of Indonesian isolates (p<0.001), but was not associated with increased chloroquine resistance after controlling for geographical location. Conclusions: In vitro susceptibility testing of P. vivax discriminates between populations with differing levels of clinical efficacy of chloroquine. The pvmdr1 polymorphism at Y976F may provide a useful tool to highlight areas of emerging chloroquine resistance, although further studies defining its clinical correlates are needed. � 2007 Suwanarusk et al.",
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    author = "Rossarin Suwanarusk and Bruce Russell and M Chavchich and Ferryanto Chalfein and Enny Kenangalem and V Kosaisavee and Prasetyorini and Kim Piera and Marion Barends and A Brockman and U Lek-uthai and Nicholas Anstey and Emiliana Tjitra and Fran{\~A}§ois Nosten and Q Cheng and Ric Price",
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    Suwanarusk, R, Russell, B, Chavchich, M, Chalfein, F, Kenangalem, E, Kosaisavee, V, Prasetyorini, Piera, K, Barends, M, Brockman, A, Lek-uthai, U, Anstey, N, Tjitra, E, Nosten, F, Cheng, Q & Price, R 2007, 'Chloroquine resistant Plasmodium vivax: in vitro characterisation and association with molecular polymorphisms', PLoS One, vol. 2, no. 10, e1089, pp. 1-9. https://doi.org/10.1371/journal.pone.0001089

    Chloroquine resistant Plasmodium vivax : in vitro characterisation and association with molecular polymorphisms. / Suwanarusk, Rossarin; Russell, Bruce; Chavchich, M; Chalfein, Ferryanto; Kenangalem, Enny; Kosaisavee, V; Prasetyorini; Piera, Kim; Barends, Marion; Brockman, A; Lek-uthai, U; Anstey, Nicholas; Tjitra, Emiliana; Nosten, François; Cheng, Q; Price, Ric.

    In: PLoS One, Vol. 2, No. 10, e1089, 31.10.2007, p. 1-9.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Chloroquine resistant Plasmodium vivax

    T2 - in vitro characterisation and association with molecular polymorphisms

    AU - Suwanarusk, Rossarin

    AU - Russell, Bruce

    AU - Chavchich, M

    AU - Chalfein, Ferryanto

    AU - Kenangalem, Enny

    AU - Kosaisavee, V

    AU - Prasetyorini, null

    AU - Piera, Kim

    AU - Barends, Marion

    AU - Brockman, A

    AU - Lek-uthai, U

    AU - Anstey, Nicholas

    AU - Tjitra, Emiliana

    AU - Nosten, François

    AU - Cheng, Q

    AU - Price, Ric

    PY - 2007/10/31

    Y1 - 2007/10/31

    N2 - Background: Treatment failure of chloroquine for P. vivax infections has reached high levels in the eastern provinces of Indonesia, however, in vitro characterization of chloroquine resistance and its associated molecular profile have yet to be determined. Methods: Using a modified schizont maturation assay we investigated the in vitro chloroquine susceptibility profile and molecular polymorphisms of P. vivax isolates collected from Papua, Indonesia, where high levels of clinical chloroquine treatment failure have been reported, and from Thailand, where chloroquine treatment is generally effective. Results: The geometric mean chloroquine IC50 for P. vivax isolates from Papua (n=145) was 312 nM [95%Cl: 237-411 nM] compared to 46.8 nM [95%Cl: 34.7-63.1 nM] from Thailand (n = 81); p<0.001. Correlating with the known clinical efficacy of the area, a cut off for chloroquine resistance was defined as 220nM, a level exceeded in 13.6% (11/81) of Thai isolates and 65% (94/145) of Papuan isolates; p<0.001. Several sequence polymorphisms in pvcrt-o and pvmdr1, and difference in pvmdr1 copy number were identified. A Y976F mutation in pvmdr1 was present in 96% (123/128) of Papuan isolates and 25% (17/69) of Thai isolates; p<0.001. Overall, the geometric mean chloroquine IC50 in isolates with the Y976F mutation was 283 nM [95%Cl: 211-379], compared to 44.5 nM [95%Cl: 31.3-63.4] in isolates with the wild type; p< 0.001. Pvmdr1 amplification occurred in 23% (15/66) of Thai isolates compared to none (0/104) of Indonesian isolates (p<0.001), but was not associated with increased chloroquine resistance after controlling for geographical location. Conclusions: In vitro susceptibility testing of P. vivax discriminates between populations with differing levels of clinical efficacy of chloroquine. The pvmdr1 polymorphism at Y976F may provide a useful tool to highlight areas of emerging chloroquine resistance, although further studies defining its clinical correlates are needed. � 2007 Suwanarusk et al.

    AB - Background: Treatment failure of chloroquine for P. vivax infections has reached high levels in the eastern provinces of Indonesia, however, in vitro characterization of chloroquine resistance and its associated molecular profile have yet to be determined. Methods: Using a modified schizont maturation assay we investigated the in vitro chloroquine susceptibility profile and molecular polymorphisms of P. vivax isolates collected from Papua, Indonesia, where high levels of clinical chloroquine treatment failure have been reported, and from Thailand, where chloroquine treatment is generally effective. Results: The geometric mean chloroquine IC50 for P. vivax isolates from Papua (n=145) was 312 nM [95%Cl: 237-411 nM] compared to 46.8 nM [95%Cl: 34.7-63.1 nM] from Thailand (n = 81); p<0.001. Correlating with the known clinical efficacy of the area, a cut off for chloroquine resistance was defined as 220nM, a level exceeded in 13.6% (11/81) of Thai isolates and 65% (94/145) of Papuan isolates; p<0.001. Several sequence polymorphisms in pvcrt-o and pvmdr1, and difference in pvmdr1 copy number were identified. A Y976F mutation in pvmdr1 was present in 96% (123/128) of Papuan isolates and 25% (17/69) of Thai isolates; p<0.001. Overall, the geometric mean chloroquine IC50 in isolates with the Y976F mutation was 283 nM [95%Cl: 211-379], compared to 44.5 nM [95%Cl: 31.3-63.4] in isolates with the wild type; p< 0.001. Pvmdr1 amplification occurred in 23% (15/66) of Thai isolates compared to none (0/104) of Indonesian isolates (p<0.001), but was not associated with increased chloroquine resistance after controlling for geographical location. Conclusions: In vitro susceptibility testing of P. vivax discriminates between populations with differing levels of clinical efficacy of chloroquine. The pvmdr1 polymorphism at Y976F may provide a useful tool to highlight areas of emerging chloroquine resistance, although further studies defining its clinical correlates are needed. � 2007 Suwanarusk et al.

    KW - chloroquine

    KW - gene product

    KW - protein pvcrt 0

    KW - protein pvmdr1

    KW - unclassified drug

    KW - antimalarial agent

    KW - protozoal protein

    KW - antibiotic resistance

    KW - antibiotic sensitivity

    KW - article

    KW - clinical effectiveness

    KW - concentration response

    KW - controlled study

    KW - correlation analysis

    KW - drug efficacy

    KW - drug treatment failure

    KW - gene amplification

    KW - gene mutation

    KW - gene number

    KW - gene sequence

    KW - genetic association

    KW - genetic polymorphism

    KW - geographic distribution

    KW - IC 50

    KW - in vitro study

    KW - Indonesia

    KW - nonhuman

    KW - nucleotide sequence

    KW - Plasmodium vivax

    KW - schizont

    KW - sensitivity analysis

    KW - Thailand

    KW - animal

    KW - biological model

    KW - chemistry

    KW - drug resistance

    KW - drug sensitivity

    KW - genetic variability

    KW - human

    KW - malaria

    KW - metabolism

    KW - parasitology

    KW - single nucleotide polymorphism

    KW - Animals

    KW - Antimalarials

    KW - Chloroquine

    KW - Drug Resistance

    KW - Humans

    KW - Inhibitory Concentration 50

    KW - Malaria, Vivax

    KW - Models, Biological

    KW - Parasitic Sensitivity Tests

    KW - Polymorphism, Genetic

    KW - Polymorphism, Single Nucleotide

    KW - Protozoan Proteins

    KW - Variation (Genetics)

    U2 - 10.1371/journal.pone.0001089

    DO - 10.1371/journal.pone.0001089

    M3 - Article

    VL - 2

    SP - 1

    EP - 9

    JO - PLoS One

    JF - PLoS One

    SN - 1932-6203

    IS - 10

    M1 - e1089

    ER -

    Suwanarusk R, Russell B, Chavchich M, Chalfein F, Kenangalem E, Kosaisavee V et al. Chloroquine resistant Plasmodium vivax: in vitro characterisation and association with molecular polymorphisms. PLoS One. 2007 Oct 31;2(10):1-9. e1089. https://doi.org/10.1371/journal.pone.0001089