Evidence of continuous use of chloroquine (cq

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)


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.


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


Aim & Objectives


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



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


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


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


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


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


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



Acknowledgements

DR. JOHNSON N. BOAMPONG

DR. NEILS B. QUASHIE

SUPERVISORS

SCHOOL OF BIOLOGICAL SCIENCES

Department of Biomedical and Forensic sciences

Health & Medicine

Evidence of continuous use of chloroquine (cq