Cargando…

Determination of anti-leishmanial drugs efficacy against Leishmania martiniquensis using a colorimetric assay

Autochthonous leishmaniasis caused by Leishmania martiniquensis cases in Thailand have dramatically increased in the recent years. L. martiniquensis infection primarily occurs in immunocompromised patients, especially AIDS patients. In Thailand, amphotericin B is the only drug available for leishman...

Descripción completa

Detalles Bibliográficos
Autores principales: Phumee, Atchara, Jariyapan, Narissara, Chusri, Saranyou, Hortiwakul, Thanaporn, Mouri, Oussama, Gay, Frederick, Limpanasithikul, Wacharee, Siriyasatien, Padet
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7153290/
https://www.ncbi.nlm.nih.gov/pubmed/32300665
http://dx.doi.org/10.1016/j.parepi.2020.e00143
_version_ 1783521622197534720
author Phumee, Atchara
Jariyapan, Narissara
Chusri, Saranyou
Hortiwakul, Thanaporn
Mouri, Oussama
Gay, Frederick
Limpanasithikul, Wacharee
Siriyasatien, Padet
author_facet Phumee, Atchara
Jariyapan, Narissara
Chusri, Saranyou
Hortiwakul, Thanaporn
Mouri, Oussama
Gay, Frederick
Limpanasithikul, Wacharee
Siriyasatien, Padet
author_sort Phumee, Atchara
collection PubMed
description Autochthonous leishmaniasis caused by Leishmania martiniquensis cases in Thailand have dramatically increased in the recent years. L. martiniquensis infection primarily occurs in immunocompromised patients, especially AIDS patients. In Thailand, amphotericin B is the only drug available for leishmaniasis treatment, and some patients relapse after amphotericin B therapy. Moreover, the efficacy of anti-leishmanial drugs against L. martiniquensis has not been evaluated to date. In this study, we determined the efficacy of various anti-leishmanial drugs against the promastigote and intracellular amastigote stages of L. martiniquensis using a colorimetric assay. Two strains (CU1 and CU1R1) were isolated from leishmaniasis HIV co-infected patient from Songkhla province, southern Thailand. The CU1 strain was isolated from the patient in 2011, and CU1R1 was isolated from the same patient in 2013, when he was diagnosed as relapse leishmaniasis. The third strain (LSCM1) used in this study has been isolated from immunocompetent patient from Lamphun province, northern Thailand. All strains were identified as L. martiniquensis by sequencing of ribosomal RNA ITS-1 and large subunit of RNA polymerase II gene. Bioassays have been conducted both with promastigote and intracellular amastigote stages of the parasite. All L. martiniquensis strains have been tested against amphotericin B, miltefosine and pentamidine to determine the efficacy of the drugs against the parasite by using a PrestoBlue. The efficacy of miltefosine and pentamidine exhibit no significant difference between each stage of L. martiniquensis among all strains. Surprisingly, the promastigote and intracellular amastigote of the CU1R1 isolate, which was isolated from a relapsed patient after amphotericin B treatment, exhibited a two-fold increased inhibitory concentration (IC50) against amphotericin B compared with other strains, and the difference was statistically significant (p < 0.05). Moreover, intracellular amastigotes isolated from CU1R1 exhibited slightly increased susceptibility to amphotericin B compared with the promastigote (p < 0.05). The result of this experiment is a scientific evident to support that in case of relapsed leishmaniasis caused by L. martiniquensis, increasing dosage of amphotericin B is essential. Moreover, this study also determined efficacy of other anti-leishmanial drugs for treatment the leishmaniasis in Thailand in case of these drugs are available in the country and the clinicians should have alternative drugs for treatment leishmaniasis in Thailand apart from amphotericin B.
format Online
Article
Text
id pubmed-7153290
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-71532902020-04-16 Determination of anti-leishmanial drugs efficacy against Leishmania martiniquensis using a colorimetric assay Phumee, Atchara Jariyapan, Narissara Chusri, Saranyou Hortiwakul, Thanaporn Mouri, Oussama Gay, Frederick Limpanasithikul, Wacharee Siriyasatien, Padet Parasite Epidemiol Control Original Research article Autochthonous leishmaniasis caused by Leishmania martiniquensis cases in Thailand have dramatically increased in the recent years. L. martiniquensis infection primarily occurs in immunocompromised patients, especially AIDS patients. In Thailand, amphotericin B is the only drug available for leishmaniasis treatment, and some patients relapse after amphotericin B therapy. Moreover, the efficacy of anti-leishmanial drugs against L. martiniquensis has not been evaluated to date. In this study, we determined the efficacy of various anti-leishmanial drugs against the promastigote and intracellular amastigote stages of L. martiniquensis using a colorimetric assay. Two strains (CU1 and CU1R1) were isolated from leishmaniasis HIV co-infected patient from Songkhla province, southern Thailand. The CU1 strain was isolated from the patient in 2011, and CU1R1 was isolated from the same patient in 2013, when he was diagnosed as relapse leishmaniasis. The third strain (LSCM1) used in this study has been isolated from immunocompetent patient from Lamphun province, northern Thailand. All strains were identified as L. martiniquensis by sequencing of ribosomal RNA ITS-1 and large subunit of RNA polymerase II gene. Bioassays have been conducted both with promastigote and intracellular amastigote stages of the parasite. All L. martiniquensis strains have been tested against amphotericin B, miltefosine and pentamidine to determine the efficacy of the drugs against the parasite by using a PrestoBlue. The efficacy of miltefosine and pentamidine exhibit no significant difference between each stage of L. martiniquensis among all strains. Surprisingly, the promastigote and intracellular amastigote of the CU1R1 isolate, which was isolated from a relapsed patient after amphotericin B treatment, exhibited a two-fold increased inhibitory concentration (IC50) against amphotericin B compared with other strains, and the difference was statistically significant (p < 0.05). Moreover, intracellular amastigotes isolated from CU1R1 exhibited slightly increased susceptibility to amphotericin B compared with the promastigote (p < 0.05). The result of this experiment is a scientific evident to support that in case of relapsed leishmaniasis caused by L. martiniquensis, increasing dosage of amphotericin B is essential. Moreover, this study also determined efficacy of other anti-leishmanial drugs for treatment the leishmaniasis in Thailand in case of these drugs are available in the country and the clinicians should have alternative drugs for treatment leishmaniasis in Thailand apart from amphotericin B. Elsevier 2020-02-19 /pmc/articles/PMC7153290/ /pubmed/32300665 http://dx.doi.org/10.1016/j.parepi.2020.e00143 Text en © 2020 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Original Research article
Phumee, Atchara
Jariyapan, Narissara
Chusri, Saranyou
Hortiwakul, Thanaporn
Mouri, Oussama
Gay, Frederick
Limpanasithikul, Wacharee
Siriyasatien, Padet
Determination of anti-leishmanial drugs efficacy against Leishmania martiniquensis using a colorimetric assay
title Determination of anti-leishmanial drugs efficacy against Leishmania martiniquensis using a colorimetric assay
title_full Determination of anti-leishmanial drugs efficacy against Leishmania martiniquensis using a colorimetric assay
title_fullStr Determination of anti-leishmanial drugs efficacy against Leishmania martiniquensis using a colorimetric assay
title_full_unstemmed Determination of anti-leishmanial drugs efficacy against Leishmania martiniquensis using a colorimetric assay
title_short Determination of anti-leishmanial drugs efficacy against Leishmania martiniquensis using a colorimetric assay
title_sort determination of anti-leishmanial drugs efficacy against leishmania martiniquensis using a colorimetric assay
topic Original Research article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7153290/
https://www.ncbi.nlm.nih.gov/pubmed/32300665
http://dx.doi.org/10.1016/j.parepi.2020.e00143
work_keys_str_mv AT phumeeatchara determinationofantileishmanialdrugsefficacyagainstleishmaniamartiniquensisusingacolorimetricassay
AT jariyapannarissara determinationofantileishmanialdrugsefficacyagainstleishmaniamartiniquensisusingacolorimetricassay
AT chusrisaranyou determinationofantileishmanialdrugsefficacyagainstleishmaniamartiniquensisusingacolorimetricassay
AT hortiwakulthanaporn determinationofantileishmanialdrugsefficacyagainstleishmaniamartiniquensisusingacolorimetricassay
AT mourioussama determinationofantileishmanialdrugsefficacyagainstleishmaniamartiniquensisusingacolorimetricassay
AT gayfrederick determinationofantileishmanialdrugsefficacyagainstleishmaniamartiniquensisusingacolorimetricassay
AT limpanasithikulwacharee determinationofantileishmanialdrugsefficacyagainstleishmaniamartiniquensisusingacolorimetricassay
AT siriyasatienpadet determinationofantileishmanialdrugsefficacyagainstleishmaniamartiniquensisusingacolorimetricassay