Double Mutation in thepfmdr1Gene Is Associated with Emergence of Chloroquine-Resistant Plasmodium falciparum Malaria in Eastern India

ABSTRACTMalaria is a major public health problem in tropical and subtropical countries, including India. This study elucidates the cause of chloroquine treatment failure (forPlasmodium falciparuminfection) before the introduction of artemisinin combination therapy. One hundred twenty-six patients were randomized to chloroquine treatment, and the therapeutic efficacy was monitored from days 1 to 28. Anin vitrosusceptibility test was performed with all isolates. Parasitic DNA was isolated, followed by PCR and restriction digestion of different codons of thepfcrtgene (codons 72 to 76) and thepfmdr1gene (N86Y, Y184F, S1034C, N1042D, and D1246Y). Finally, sequencing was done to confirm the mutations. Forty-three (34.13%) early treatment failure cases and 16 (12.69%) late treatment failure cases were observed after chloroquine treatment.In vitrochloroquine resistance was found in 103 isolates (81.75%). Twenty-six (60.47%) early treatment failure cases and 6 (37.5%) late treatment failure cases were associated with the CVMNK-YYSNYallele (the underlined amino acids are those that were mutated). Moreover, the CVIEK-YYSNYallele was found in 8 early treatment failure (18.60%) and 2 late treatment failure (12.5%) cases. The presence of the wild-typepfcrt(CVMNK) andpfmdr1(YYSNY) double mutant allele in chloroquine-nonresponsive cases was quite uncommon.In vivochloroquine treatment failure andin vitrochloroquine resistance were strongly correlated with the CVMNK-YYSNYand CVIEK-YYSNYhaplotypes (P< 0.01).

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Spread of multidrug-resistant Plasmodium falciparum malaria and predictors of treatment failures in Vietnam

10.21203/rs.2.18720/v1 ◽

2019 ◽

Author(s):

Minh Cuong Duong ◽

Oanh Kieu Nguyet Pham ◽

Phong Thanh Nguyen ◽

Van Vinh Chau Nguyen ◽

Phu Hoan Nguyen

Keyword(s):

Plasmodium Falciparum ◽

Treatment Failure ◽

Severe Malaria ◽

Falciparum Malaria ◽

Early Treatment ◽

Molecular Technique ◽

Drug Resistant ◽

Early Treatment Failure ◽

Central Highland ◽

Late Treatment

Abstract Background Drug-resistant falciparum malaria is an increasing public health burden. We examined the magnitude of Plasmodium falciparum infection and the patterns and predictors of treatment failure in Vietnam. Methods Medical records of all 443 patients with malaria infection admitted to the Hospital for Tropical Diseases between January 2015 and December 2018 were used to extract information on demographics, risk factors, symptoms, laboratory tests, treatment, and outcome. Results More than half (59.8%, 95%CI 55.1%-64.4%) of patient acquired P. falciparum infection of whom 21.9% (95%CI 17.1%-27.4%) had severe malaria, while 10.2% (95%CI 6.8%-14.5%) and 19.2% (95%CI 14.7%-24.5%) developed early treatment failure (ETF) and late treatment failure (LTF) respectively. ETF was 6.8% among patients referred from Binh Phuoc province and Central Highland, 11.3% from other areas in Vietnam, and 6.9% from Africa. LTF was 16.2% among patients from Binh Phuoc province and Central Highland, 22.6% from other areas in Vietnam, and 27.6% from Africa. Most patients (98.5%) recovered completely. Having severe malaria was a predictor of ETF (AOR 4.42, 95%CI 1.85-10.61, P = 0.001). No predictor of LTF was identified.Conclusion P. falciparum remains the prevalent malaria parasite. Despite low mortality rate, severe malaria is not rare and is a significant predictor of ETF. Parenteral artesunate and an oral partner drug should be concurrently used for severe malaria to reduce the risk for ETF. The study alerts the possibility of drug-resistant malaria in Africa and other areas in Vietnam which are known as nonendemic areas of antimalarial drug resistance. A more comprehensive study using molecular technique in these regions is required to completely understand the magnitude of drug-resistant malaria and to design appropriate control strategies. Key words: Plasmodium falciparum; severe malaria; early treatment failure; late treatment failure; Vietnam

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Early treatment failure during treatment of Plasmodium falciparum malaria with atovaquone-proguanil in the Republic of Ivory Coast

Malaria Journal ◽

10.1186/1475-2875-11-146 ◽

2012 ◽

Vol 11 (1) ◽

Cited By ~ 8

Author(s):

Nathalie Wurtz ◽

Aurélie Pascual ◽

Adeline Marin-Jauffre ◽

Housem Bouchiba ◽

Nicolas Benoit ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Treatment Failure ◽

Falciparum Malaria ◽

Early Treatment ◽

Ivory Coast ◽

Plasmodium Falciparum Malaria ◽

Early Treatment Failure ◽

The Republic

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Pfcrt mutant haplotypes may not correspond with chloroquine resistance

The Journal of Infection in Developing Countries ◽

10.3855/jidc.3398 ◽

2014 ◽

Vol 8 (06) ◽

pp. 768-773 ◽

Cited By ~ 6

Author(s):

Diganta Goswami ◽

Sunil Dhiman ◽

Bipul Rabha ◽

Dinesh Kumar ◽

Indra Baruah ◽

...

Keyword(s):

Treatment Failure ◽

Treatment Success ◽

Chloroquine Resistance ◽

Pfcrt Gene ◽

Kaplan Meier ◽

Chloroquine Treatment ◽

Pcr Rflp ◽

Pcr Products ◽

Late Treatment

Introduction: Chloroquine resistance in Plasmodium falciparum is associated with mutations in pfcrt and pfmdr1 genes. The frequency distribution of pfcrt K76T and pfmdr1 N86Y mutations and their association with chloroquine susceptibility was studied in an endemic area along the Indo-Bangladesh border. Methodology: A single-arm prospective study of clinical and parasitological responses in P. falciparum malaria patients to chloroquine was conducted in vivo. PCR-RFLP assay was used to detect pfcrt K76T and pfmdr1 N86Y mutations in P. falciparum. The PCR products of pfcrt gene were sequenced, translated and aligned for haplotyping. Results: Out of 63 cases, 44 (69.8%) responded adequately to chloroquine treatment. Pfcrt K76T mutation was recorded in 100% of the treatment failure cases, whereas pfmdr1 N86Y mutation was found in 52.6% of the cases only. Early treatment failure (84.2%) occurred more frequently than late treatment failure (15.8%). Kaplan–Meier survival analysis showed that the probability estimate for treatment success after 7 and 15 days was 0.84 (95% CI = 0.72-0.92) and 0.70 (95% CI = 0.57-0.80), respectively. Sequence analysis of 72 to 76 pfcrt gene codons revealed the presence of two mutant (CVMNT, CVIET) and two wild (CVMNK, CVIEK) haplotypes. The mutant CVIET haplotype was predominantly distributed (42.1%). Conclusions: The presence of mutations in pfcrt K76T and pfmdr1 N86Y genes is not sufficient to explain the therapeutic efficacy of chloroquine to P. falciparum. Study suggests that pfcrt K76T mutant haplotypes are widely distributed and are spreading diligently, which needs to be taken into account in devising an antimalarial policy.

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Efikasi Obat Kloroquine, Kina, Artesunate-SP, Artesunate-Amodiaquine, Artesunate-Lumafentrin pada Anak Malaria Falciparum di BLU RSUP Prof. Dr. RD. Kandou Manado

Sari Pediatri ◽

10.14238/sp10.6.2009.417-23 ◽

2016 ◽

Vol 10 (6) ◽

pp. 417

Author(s):

Suryadi Nicolaas Napoleon Tatura

Keyword(s):

Plasmodium Falciparum ◽

Treatment Failure ◽

Early Treatment ◽

Early Treatment Failure ◽

Negative Rate ◽

Kaplan Meier

Latar belakang. Malaria falciparum masih merupakan salah satu penyebab utama kematian dan kesakitan pada anak-anak dan orang dewasa di negara-negara tropis. Di Indonesia, dilaporkan Plasmodium falciparum telsh resisten terhadap obat – obat anti malaria, terutama kemungkinan terjadi early treatment failure (ETF).Tujuan. Untuk mengetahui efikasi dan early treatment failure (ETF) obat anti malaria (OAM) yaitu kloroquin (CQ), kina, artesunat-SP(AS-SP), dan artesunate-amodiaquin(AS-AQ) serta artesunate- lumafentrin(AS-L) pada anak dengan malaria falciparum.Metode. Penelitian deskriptif retrospektif� �� . Populasi adalah bayi dan anak usia 1 bulan-14 tahun yang terdiagnosis dengan malaria falciparum dan mendapat OAM. Data diperoleh dari rekam medis Bagian Anak BLU RSUP Prof Dr. RD. Kandou Manado sejak Maret 2007 sampai Maret 2009. Data dikelompokkan berdasarkan jenis obat anti malaria yang digunakan oleh pasien selama 3 hari (3x24jam), kemudian dinilai waktu bebas parasit dalam darah pasien serta ETF. Hitung parasit menggunakan metode semikuantitatif. Data dianalis dengan metode Kaplan-Meier menggunakan SPSS 17.Hasil. Efikasi obat anti malaria dalam 3 hari sebagai berikut, AS-L 100%, AS-SP 100%, AS-AQ 97%, CQ 85%, dan kina 81%. Terdapat ETF obat kina 19%, CQ 15% dan AS-AQ 3%. Parasite negative rate dalam 24 jam AS-SP 0,6, AS-L 0,6, AS-AQ 0,89, Kina 0,35 dan CQ 0,54.Kesimpulan. Artesunate-lumafentrin dan artesunate-SP merupakan obat anti malaria falciparum pilihan. Artesunate-amodiaquine sangat baik menurunkan angka parasit dalam 24 jam I. Telah terjadi ETF pada kloroquine, kina dan arteunate-amodiaquine.

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Chloroquine treatment for uncomplicated childhood malaria in an area with drug resistance: early treatment failure aggravates anaemia

Transactions of the Royal Society of Tropical Medicine and Hygiene ◽

10.1016/s0035-9203(98)90913-0 ◽

1998 ◽

Vol 92 (5) ◽

pp. 556-560 ◽

Cited By ~ 32

Author(s):

H. Ekvall ◽

Z. Premji ◽

A. Björkman

Keyword(s):

Drug Resistance ◽

Treatment Failure ◽

Early Treatment ◽

Early Treatment Failure ◽

Chloroquine Treatment ◽

Childhood Malaria

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Micro in vitro test for chloroquine resistance of Plasmodium, falciparum: dicrepancies attributable to batch variation of plates

Transactions of the Royal Society of Tropical Medicine and Hygiene ◽

10.1016/0035-9203(87)90183-0 ◽

1987 ◽

Vol 81 (3) ◽

pp. 513 ◽

Cited By ~ 1

Author(s):

S. Sinha ◽

A. Gajanana

Keyword(s):

Plasmodium Falciparum ◽

Chloroquine Resistance ◽

In Vitro Test ◽

Vitro Test

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Piperaquine resistant Cambodian Plasmodium falciparum clinical isolates: in vitro genotypic and phenotypic characterization

10.21203/rs.3.rs-22444/v3 ◽

2020 ◽

Author(s):

Nonlawat Boonyalai ◽

Brian A Vesely ◽

Chatchadaporn Thamnurak ◽

Chantida Praditpol ◽

Watcharintorn Fagnark ◽

...

Keyword(s):

Drug Resistance ◽

Plasmodium Falciparum ◽

Copy Number ◽

Drug Susceptibility ◽

Antagonistic Activity ◽

Drug Combinations ◽

Chloroquine Resistance ◽

Phenotypic Characterization ◽

Field Isolates

Abstract Background High rates of dihydroartemisinin-piperaquine (DHA-PPQ) treatment failures have been documented for uncomplicated Plasmodium falciparum in Cambodia. The genetic markers plasmepsin 2 ( pfpm2 ), exonuclease ( pfexo ) and chloroquine resistance transporter ( pfcrt ) genes are associated with PPQ resistance and are used for monitoring the prevalence of drug resistance and guiding malaria drug treatment policy.Methods To examine the relative contribution of each marker to PPQ resistance, in vitro culture and the PPQ survival assay were performed on seventeen P. falciparum isolates from northern Cambodia, and the presence of E415G-Exo and pfcrt mutations (T93S, H97Y, F145I, I218F, M343L, C350R, and G353V) as well as pfpm2 copy number polymorphisms were determined. Parasites were then cloned by limiting dilution and the cloned parasites were tested for drug susceptibility. Isobolographic analysis of several drug combinations for standard clones and newly cloned P. falciparum Cambodian isolates was also determined.Results The characterization of culture-adapted isolates revealed that the presence of novel pfcrt mutations (T93S, H97Y, F145I, and I218F) with E415G-Exo mutation can confer PPQ-resistance, in the absence of pfpm2 amplification. In vitro testing of PPQ resistant parasites demonstrated a bimodal dose-response, the existence of a swollen digestive vacuole phenotype, and an increased susceptibility to quinine, chloroquine, mefloquine and lumefantrine. To further characterize drug sensitivity, parental parasites were cloned in which a clonal line, 14-B5, was identified as sensitive to artemisinin and piperaquine, but resistant to chloroquine. Assessment of the clone against a panel of drug combinations revealed antagonistic activity for six different drug combinations. However, mefloquine-proguanil and atovoquone-proguanil combinations revealed synergistic antimalarial activity.Conclusions Surveillance for PPQ resistance in regions relying on DHA-PPQ as the first-line treatment is dependent on the monitoring of molecular markers of drug resistance. P. falciparum harbouring novel pfcrt mutations with E415G-exo mutations displayed PPQ resistant phenotype. The presence of pfpm2 amplification was not required to render parasites PPQ resistant suggesting that the increase in pfpm2 copy number alone is not the sole modulator of PPQ resistance. Genetic background of circulating field isolates appear to play a role in drug susceptibility and biological responses induced by drug combinations. The use of latest field isolates may be necessary for assessment of relevant drug combinations against P. falciparum strains and when down-selecting novel drug candidates.

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Drug sensitivity of Plasmodium falciparum field isolates to selected antimalarial drugs in Ghana using the in-vitro DAPI assay

10.21203/rs.3.rs-16372/v1 ◽

2020 ◽

Author(s):

Michael Fokuo Ofori ◽

Emma E. Kploanyi ◽

Benedicta A. Mensah ◽

Emmanuel K. Dickson ◽

Eric Kyei Baafour ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Drug Sensitivity ◽

Geometric Mean ◽

Antimalarial Drugs ◽

Chloroquine Resistance ◽

Cape Coast ◽

African Region ◽

Cross Sectional ◽

Ecological Zones

Abstract Background: Malaria continues to be a major health issue globally with nine out of ten cases reported in Africa. Although the current artemisinin derived combination therapies in Ghana are still efficacious against the Plasmodium falciparum parasite, compounding evidence of artemisinin and amodiaquine resistance in the African region establish the need for a full, up-to-date understanding and monitoring of antimalarial resistance to provide evidence for planning control strategies.Methods: The study was cross-sectional and was conducted during the peak transmission seasons of 2015, 2016, and 2017 in two study sites located in different ecological zones of Ghana involving children aged 0.5-14 years presenting with symptomatic uncomplicated Plasmodium falciparum (Pf) malaria with parasitaemia greater than 1000 parasites/µl of blood. Using in vitro 4-,6-diamidino-2-phenylindole (DAPI) drug sensitivity assays, 328 Pf parasites collected were used to investigate susceptibility to five selected antimalarial drugs: chloroquine, amodiaquine, dihydroartemisinin, artesunate and mefloquine.Results: The geometric mean B (GMIC50) of five drugs against the parasites collected from Cape Coast were 9.6, 23.6, 9.1, 3.5 and 8.1nM for chloroquine, amodiaquine, artemisinin, artesunate, and mefloquine respectively in 2015. There was a 2 fold increase in the GMIC50 levels of all the drugs against the isolates collected in 2016 as compared to the 2015 data from Cape Coast .The a of the five drugs against the parasites collected from Cape Coast were significantly higher than those isolates collected from Begoro in 2016 and 2017 (P<0.001) . The chloroquine resistance ranged between 1.9% and 9.1% among isolates collected from Cape Coast but remained 0% in Begoro over the period. High amodiaquine resistance levels were recorded at both sites whilst that of artesunate resistance ranged between 4 and 10% over the study period.Conclusions: The study has assessed the antimalarial drug sensitivities of Ghanaian Pf isolates collected over 3 consecutive years. The parasites showed variable resistance levels to all the drugs used over the period. The study has demonstrated the continual return of chloroquine-sensitive parasites. The in vitro DAPI assay is a useful method for monitoring individual drugs used in combinations in Ghana for the generation of data on their sensitivities over time.

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Mycobacterium abscessus Complex Infections: A Retrospective Cohort Study

Open Forum Infectious Diseases ◽

10.1093/ofid/ofy022 ◽

2018 ◽

Vol 5 (2) ◽

Cited By ~ 18

Author(s):

Maroun Sfeir ◽

Marissa Walsh ◽

Rossana Rosa ◽

Laura Aragon ◽

Sze Yan Liu ◽

...

Keyword(s):

Risk Factors ◽

Cohort Study ◽

Renal Disease ◽

Treatment Failure ◽

Retrospective Cohort Study ◽

Early Treatment ◽

Retrospective Cohort ◽

Positive Culture ◽

Mycobacterium Abscessus ◽

Early Treatment Failure

Abstract Background Infections caused by Mycobacterium abscessus group strains are usually resistant to multiple antimicrobials and challenging to treat worldwide. We describe the risk factors, treatment, and clinical outcomes of patients in 2 large academic medical centers in the United States. Methods A retrospective cohort study of hospitalized adults with a positive culture for M. abscessus in Miami, Florida (January 1, 2011, to December 31, 2014). Demographics, comorbidities, the source of infection, antimicrobial susceptibilities, and clinical outcomes were analyzed. Early treatment failure was defined as death and/or infection relapse characterized either by persistent positive culture for M. abscessus within 12 weeks of treatment initiation and/or lack of radiographic improvement. Results One hundred eight patients were analyzed. The mean age was 50.81 ± 21.03 years, 57 (52.8%) were females, and 41 (38%) Hispanics. Eleven (10.2%) had end-stage renal disease, 34 (31.5%) were on immunosuppressive therapy, and 40% had chronic lung disease. Fifty-nine organisms (54.6%) were isolated in respiratory sources, 21 (19.4%) in blood, 10 (9.2%) skin and soft tissue, and 9 (8.3%) intra-abdominal. Antimicrobial susceptibility reports were available for 64 (59.3%) of the patients. Most of the isolates were susceptible to clarithromycin, amikacin, and tigecycline (93.8%, 93.8%, and 89.1%, respectively). None of the isolates were susceptible to trimethoprim/sulfamethoxazole, and only 1 (1.6%) was susceptible to ciprofloxacin. Thirty-six (33.3%) patients early failed treatment; of those, 17 (15.7%) died while hospitalized. On multivariate analysis, risk factors significantly associated with early treatment failure were disseminated infection (odds ratio [OR], 11.79; 95% confidence interval [CI], 1.53–81.69; P = .04), acute kidney injury (OR, 6.55; 95% CI, 2.4–31.25; P = .018), organ transplantation (OR, 2.37; 95% CI, 2.7–23.1; P = .005), immunosuppressive therapy (OR, 2.81; 95% CI, 1.6–21.4; P = .002), intravenous amikacin treatment (OR, 4.1; 95% CI, 0.9–21; P = .04), clarithromycin resistance (OR,79.5; 95% CI, 6.2–3717.1, P < .001), and presence of prosthetic device (OR, 5.43; 95% CI, 1.57–18.81; P = .008). Receiving macrolide treatment was found to be protective against early treatment failure (OR, 0.13; 95% CI, 0.002–1.8; P = .04). Conclusions Our cohort of 108 M. abscessus complex isolates in Miami, Florida, showed an in-hospital mortality of 15.7%. Most infections were respiratory. Clarithromycin and amikacin were the most likely agents to be susceptible in vitro. Resistance to fluoroquinolone and trimethoprim/sulfamethoxazole was highly common. Macrolide resistance, immunosuppression, and renal disease were significantly associated with early treatment failure.

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