evidence of continuous use of chloroquine (cq
TRANSCRIPT
Evidence of continuous use of Chloroquine (CQ) in Ghana after seven years of its ban, its consequences on CQ resistant markers
ASARE, KUMI KWAME
Department of Biomedical and Forensic sciences, University of Cape Coast
“you read about it in the news but you don't believe it you'll only know about it when the man in the long black coat knocks on your door 'cause you're his next victim”…Lucky Dube
Introduction
K. K. Asare, et al., 2
CQ resistance
Late 1950s and early 1960s
In 1965 CQ resistance had been alleged by Schwendler (Beausoleil, 1967).CQ resistant P. falciparum were isolated in 1986 (Neequaye, 1986)
K. K. Asare, et al., 3
In 2001, CQ resistance P. falciparum in Ghana was reported to be around 88% (Evans et al., 2005)
In 2005,MoH finally implemented ACTs
Despite the replacement anecdotal evidence suggests that CQ is still in used in many communities in the country.
K. K. Asare, et al., 4
The continuous use of the former drug would result in development of stable mutations in:
I. Pfcrt gene
II. Pfmdr-1 geneWhich can lead to cross-resistance to other
antimalarial drugs such as:
1. Amodiaquine
2. Artemisinin
3. Quinine
K. K. Asare, et al., 5
Aim & Objectives
K. K. Asare, et al., 6
Methodology
Solomon-Saker method
I.0.05%TBPEE-in-chloroform
Molecular method
I.Chelex dna extraction
II.Nested PCR methods
III.RFLP
Statistical analysis: SPSS-Chi-square at 95% CI
K. K. Asare, et al.,
STUDY SITES
SUBJECT SELECTION
MALARIA INFECTED PATIENTS
NON-MALARIA INFECTED PATIENTS
SUBJECTS WHO WERE NOT ENROLLED
ENROLLED SUBJECTS
SAMPLE TAKEN
BLOOD SAMPLE URINE SAMPLE
MYSTERY BUYING METHOD
SALES OF CQ BY COMMUNITY PHARMACY & CHEMICAL SHOPS
1. HAEMATOLOGICAL ANALYSIS 2. MOLECULAR ANALYSIS
1. URINALYSIS 2. URINE CQ ANALYSIS
1. ELMINA HEALTH CENTRE 2. CAPE COAST DISTRICT HOSPITAL 3. TWIFO PRASO DISTRICT HOSPITAL 4. ST. FRANCIS XAVIER HOSPITAL, ASSIN
FOSO
ETHICAL CLEARANCE:
1. GHSERC 2. UCCIRB
QUESTIONNAIRE ADMINISTRATION
1
2
7
RESULTS AND DISCUSSION
K. K. Asare, et al., 8
K. K. Asare, et al.,
Fig.1: Samples of chloroquine injections
1. They were mainly manufactured at India & China
2. About 20% of CQ injections obtained did not have labels
FIG. 1
9
K. K. Asare, et al.,
Fig.2: controls samples fortified with CQ and patients samples that contained CQ Cross-reactivity test were performed Contrary to Malawi Asia & South-America case, Ghana has maintained high CQR due to continuous
CQ usage
FIG 2
10
K. K. Asare, et al.,
Fig. 3: Apo 1 RFLP at K76T in pfcrt
Persistent CQ use subject the parasite to drug pressure
Development of stable T-76 mutation & other set of mutations in pfcrt gene
200
100
234
FIG. 3 FIG. 4
11
K. K. Asare, et al.,
Fig. 5: Dpn II RFLP at D1246Y in pfmdr-1
Mutations in pfmdr-1 is synergetic to pfcrt in CQR mechanisms
Combination of haplotype mutations in pfmdr-1 modulates CQR
800
400
100
FIG. 5 FIG.6
12
• 86 Y & 184 F mutations play a role in CQR
• Studies from South-America, Asia & Africa have shown that 86 Y & 184 F confer resistance to QN, MFQ & Halofantrine
• Also modulate parasite sensitivity to artemisinin drug
K. K. Asare, et al., 13
Initial studies reported association between 86 Y in pfmdr-1 & CQR
However, several field studies showed inconsistency
Again, transfection studies showed that replacement of 86 Y with 86 N in pfmdr-1 decrease CQR from high to moderate levels
K. K. Asare, et al., 14
CONCLUSION & RECOMMENDATION
The study has revealed continuous use of CQ in the country Increase of point mutations in pfmdr-1 gene These mutations can cause cross-resistance to current anti-
malarial drugs
We recommend:
a. Stringent regulations
b. Further studies:
1. To investigate CQ importation
2. Drug sensitivity and pharmacokinetics studies to validate our findings
K. K. Asare, et al., 15
K. K. Asare, et al., 16
Acknowledgements
DR. JOHNSON N. BOAMPONG
DR. NEILS B. QUASHIE
SUPERVISORS
SCHOOL OF BIOLOGICAL SCIENCES
Department of Biomedical and Forensic sciences