Surveillance on drug resistance in tuberculosis C N Paramasivan Tuberculosis Research centre (ICMR)
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Transcript of Surveillance on drug resistance in tuberculosis C N Paramasivan Tuberculosis Research centre (ICMR)
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Role of TRC in DRS for India & SEARRole of TRC in DRS for India & SEAR SNRL and ref. Lab of the WHO NRL for India
Renders assistance for the following: Preparation of generic protocol Developing laboratories for culture & DST Preparation of manuals and SOPs Training of laboratory personnel Instituting uniform methods for DST Ensuring quality thro’ QAP Supply of standard strains, drugs & reagents Periodic site visits
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Earlier reports on combined resistance Earlier reports on combined resistance from India and their limitationsfrom India and their limitations
Case selection
Sample size
Methodology
Source of drugs
Definition of resistance
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MDR TB in SEARMDR TB in SEAR
4
1.3
0.9
15.5
20.5
20.2
0 5 10 15 20 25
Treated
New
Thailand (2000 – 02)
172
1505
Nepal (2000 – 02)
177
755
Myanmar (2003 – 04)
166
733
Percentage
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Prevalence of primary DRPrevalence of primary DR TRC Studies 1956 - 2001TRC Studies 1956 - 2001
Prevalence of primary DRPrevalence of primary DR TRC Studies 1956 - 2001TRC Studies 1956 - 2001
0
2
4
6
8
10
12
14
16
18
YEARS
H
S
SH
R/H
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Prevalence of Primary Drug Resistance Prevalence of Primary Drug Resistance TRC Studies – (1974 -2001)TRC Studies – (1974 -2001)
0
5
10
15
20
XII 74-77
XIII 77-80
XIV80-85
XV 85-86
XVI 86-90
XVII90-95
XVIII95-98
XIX 98-00
XXI2001
per
cen
tag
e
MDR
RIF
STREP
INH
After the introduction of rifampicin in Controlled Clinical Trail at TRC
*
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DRS sites of India DRS sites of India (1985-2003)(1985-2003)
North Arcot – 1985-89 (2%), 1989-90 (1.7%), 1999 (3%)
Pondicherry - 1985 (0.9%)
Tamil Nadu - 1995 (3.3%)
AFMS – 1995 -1999 (2.7%)
Bangalore - 2002 (2.2%)
Mysore - 2001 (1.2%)
Raichur - 1989 (3.2%), 1999 (2.5%)
Wardha - 2001 (0.5%)
Jabalpur - 2002 (1%)
Mayurbhanj - 2002 (0.7%)
Hoogli - 2003 (3%)
Population covered = 8.1%
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Level of MDR in ‘New’ in different Level of MDR in ‘New’ in different sites in Indiasites in India
3.3%
3.2%
3.0%
3.0%
2.7%
2.5%
2.2%
2.0%
1.7%
1.2%
1.0%
0.9%
0.7%
0.5%
0.0% 1.0% 2.0% 3.0% 4.0% 5.0% 6.0% 7.0% 8.0% 9.0% 10.0%
TAMILNADU (389) (1995)
RAICHUR (244) (1989)
HOOGLY (350) (2003)
NORTH ARCOT (282) (1999)
AFMS (2562) (1995-99)
RAICHUR (278) (1999)
BANGALORE (366) (2002)
NORTH ARCOT (2779) (1985-89)
NORTH ARCOT (350) (1989-90)
MYSORE (203) (2001)
JABALPUR (273) (2002)
PONDICHERRY (2127) (1985)
MAYURBHANJ (343) (2002)
WARDHA (197) (2001)
Percent MDR
(population covered 8.1%)(population covered 8.1%)
TRCNTI
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DRS in MDP area
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85
73
15
27
10 13
1.7 2.20
10
20
30
40
50
60
70
80
90
U n T re a te d
fully sens.
Any Res.
Any H
HR
Drug resistance among Drug resistance among newly diagnosed casesnewly diagnosed cases
MDP area N=1603 Bangalore city N=271
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59 60
41 4037
27
12 13
0
10
20
30
40
50
60
70
R e tre a tme n t c a se s
Fully densitive
Any Res.
Any H
HR
Drug susceptibility among previously Drug susceptibility among previously treated casestreated cases
MDP area N=443 Bangalore city N=226
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9.8 9.3 9.7
2.7
41.8
34.0 35.037.6
21.7
9.8
16.0
0.9 1.1 1.03.5
41.3
10.812.611.2
8.0
0
5
10
15
20
25
30
35
40
45
1999 2000 2001 2002 2003
per
cen
tag
e
H (New)
HR (New)
H (Re Rx)
HR (Re Rx)
Drug resistance trend in MDP areaDrug resistance trend in MDP area
New 144 326 367 389 371
Re Rx. 46 98 100 103 93
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MDR TB MDR TB (Gujarat, India Jan 2000 – Aug 2001)(Gujarat, India Jan 2000 – Aug 2001)
60
35
0
10
20
30
40
50
60
70
80
90
100
All patients Res. cases
N=822
N=482
Shah AR et al. Int J tuberc Lung Dis, 2003; 6(12): 1098.
3.9 %
12.4 %
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Drug Resistance in Patients With Drug Resistance in Patients With HIV / TB in South IndiaHIV / TB in South India
62.2
13.6
36.8
13.2
27
4.2
13.5
84.4
0
10
20
30
40
50
60
70
80
90
% o
f P
ati
en
ts
suseptible to all Any resis.
Res. To H MDR
New cases-167 Treated cases-37
Swaminathan S et al IJTLD 2004
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Drug Resistance pattern of Drug Resistance pattern of referred samples 2001-04 referred samples 2001-04
Susc. 32.5%
Res. 1 or more 67.5%
(n 2816 patients)(n 2816 patients)
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Drug Resistance pattern of Drug Resistance pattern of referred samples 2001-04 referred samples 2001-04
(n 2816 patients)(n 2816 patients)
43.2
67.5
1
29.9
6.5
24.6
9.7
53.2
0
20
40
60
80
100
S H R E K Th Ofl. HR
Drugs
( % )
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Drug Resistance pattern of referred samples Drug Resistance pattern of referred samples
2001-042001-04 (n 2816 patients)(n 2816 patients)
29 6
2011
23
30 36
40
337
1214
26
16
68
49 44
14
40
0
10
20
30
40
50
60
70
80
90
100
HR HRS HRE HRSE Any HRRes.
per
cen
tag
e
Column 4
O
Eth
K
N 355
385 176 582 1498
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Few earlier studies on ADR in India
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Level of MDR in ‘New’ in different Level of MDR in ‘New’ in different sites in Indiasites in India
9.6
15
33.7
28.0
20.3
30.0
35.2
11.0
13.5
17.0
0 5 10 15 20 25 30 35 40
WARDHA (302) (1982-89)
Gujarat (1267) (1983-86)
Delhi (87) (1990-91)
Bombay (521) (1991-95)
T.Nadu (162) (1995)
Lucknow (183) (2001)
Gujarat (822) (2002)
NA Dt (560) (1986-88)
NA (S) (37) (1989-90)
Raichur (S) (111) (1989)
Percent MDR
(population covered 8.1%)(population covered 8.1%)
TRCOthers
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Year 2005 DRS sites of India Year 2005 DRS sites of India
Maharashtra, 102.8 millions (9.4%), I Qrt 2005
Gujarat, 53.8 millions (4.9%), I Qrt 2005
Orissa, 38.2 millions (3.5%), II Qrt 2005
Andhra Pradesh, 78.7 millions (7.2%), II Qrt 2005
Population being covered in 2005 = 25%
Resurvey – Tamilnadu DRS-Sikkim,2005-06
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TRC Studies on Newer drugs and TRC Studies on Newer drugs and defining resistancedefining resistance
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Studies carried out at TRCStudies carried out at TRC
DEFINITION OF RESISITANCE TO RIFAMPICINDEFINITION OF RESISITANCE TO RIFAMPICIN
MIC MIC : 128 : 128µg/ml. µg/ml.
PST (1% or more) : 40 µg/ml PST (1% or more) : 40 µg/ml
BACTEC BACTEC : 2 µg/ml : 2 µg/ml
Indian J. Med. Res. 2001, 114, 187-191.Indian J. Med. Res. 2001, 114, 187-191.
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Studies carried out at TRC Studies carried out at TRC
In vitro activity of capreomycin and ciprofloxacin against S.Indian isolates of M.tb Indian J Tuberculosis 1993; 40: 21-25
In vitro activity of ciprofloxacin and ofloxacin against S.Indian isolates of M.tb Indian J Tuberculosis 1994; 41: 87-90
MIC of Lomefloxacin and Minocycline Against Drug-
Sensitive & resistant Isolates of M.tuberculosis Compared on L-J and 7H11 Media Int J Leprosy 1997; 65: 375-378
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Studies carried out at TRCStudies carried out at TRC
Evaluation of various methods of susceptibility to ofloxacin in strains of M.tb Indian J Med Res 1999; 110: 186-189 Evaluation of bactericidal action of ofloxacin and sulbactam/ampicillin alone & in combination with R & H on M.tb invitro Antimicrob Agents Chemother 1996; 40: 2296-2299
A multi centre study of the early bactericidal activity of anti- tuberculosis drugs
J Antimicrobial Chemother 2000; 45: 859-870
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Recent TRC studies on newer QuinolonesRecent TRC studies on newer Quinolones
1. Bactericidal action of Gatifloxacin, Rifamicin and isoniazid on Logarithmic –
and Stationary – Phase Cultures of Mycobacterium tuberculosis. Antimicrob.
Agents Chemother.2005, 49:627 – 631
2. Moxifloxacin and Gatifloxacin in a new acid model of persistent
M.tuberculosis. Antimicrob. Agents Chemother.2005
3. In vitro activity of fluoroquinlones against M.tuberculosis. J.
Chemotherapy.April,2005 (Accepted)
4. In vitro definitions of MIC of gati and moxifloxacin by different test methods.
FEMS Microbiology.2005
5. Bactericidal action of Moxifloxacin, Rifampicin and Isoniazid on Logarithmic –
and Stationary phase cultures of M.tuberculosis. J Antimicro.Agents and
Chemother. (2005 Communicated)
6. Analysis of Fluoroquinolone resistance in clinical isolates of M.tuberculosis
from India. J. Clinical Microbiology,2005 ( Communicated)
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In vitro definition of resistance to gatifloxacin & Moxifloxacin
No. of strains : 50 (Sens. 30; Res. 20) Methods used : Abs.conc. - LJ PST - LJ, 7H11 & BACTECRESULTSRESULTS
MIC of GATI LJ : 1 μg ml
Critical conc. LJ & 7H11 : 0.5 μg/ml
BACTEC : 0.25 μg/ml
MIC of MOXI LJ : 1 μg /ml
Critical conc. LJ : 1 μg/ml
7H11 & BACTEC : 0. 5 μg/ml
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No of strains : 55 (oflox-Res 33; Susc 22)
Method of testing : MIC
Drugs tested : Spar, Oflo, Cipro, Lome,moxi & Gati
Media used : LJ & 7H11
RESULT : Fluoroquinolones exhibited cross resistance
at different levels.
MIC of quinolones were in the order of GTFX = MOXI >
SPFX > OFLX > CFLX > LMFX
TRC Study
J.Chemother,2005
Determination of MIC & Cross Determination of MIC & Cross resistance in resistance in M.tbM.tb
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TRC study findingsTRC study findings
In vitroIn vitro MIC studies MIC studies
Quinolones showed low and similar MIC on both drug sens & resist. population of M.tb
Cipro showed higher mean MIC than Ofloxacin
Almost 100% cross resistance was seen
Ofloxacin MICs were lower than other quinolones tested
PST on LJ showed 2mg/l as a criterion of resistance for Ofloxacin
Absolute Concentration Method (Ofloxacin) :8mg/l
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TRC study findingsTRC study findings
In vitroIn vitro simulation experiment with ofloxacin simulation experiment with ofloxacin
Showed high EBA either alone or in combinations on
exponential growth
Expect high bactericidal activity in the early phase of the
Rx
Comparatively low level of SA against stationary phase
growth
However, it enhanced activity in combination with H, R &
HR
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Definition of resistance to QuinolonesDefinition of resistance to Quinolones
OFLO : NCCLS 2.0 µg/ml (7H10 & 7H11) TRC 8 µg/ml ACM (LJ) 2 µg/ml PST (LJ)
GATI : TRC 1 µg/ml – LJ; 0.5 & 0.25 (7H11 & BACTEC)
MOXI : TRC 1 µg/ml – (LJ); 0.5 (7H 11 & BACTEC)
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Standardisation of DST to newer drugsStandardisation of DST to newer drugs
• NCCLS (2002) Guidelines :
7H10 & 7H 11: Capreo, Eth, Kan, Oflo, PAS, RBU & Strep
BACTEC : PZA
• Canetti etal (1969)& Various TRC Publications LJ : INH, Capreo, Amikacin,Rif,RBU, Kan, Eth, Cyclo
• TRC: Lomi, Cipro,Oflo,Gati & Moxi
• Developing SOP for country’s requirement
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Type Geometric mean LJ 7H11 Sensitive(46) 63.97 26.73(SHR)
Resistant (46) 65.92 23.82(SHR/HR) Total (92) 65.01 25.23
MIC of S/A against sensitive and MIC of S/A against sensitive and resistant isolates of M.tuberculosisresistant isolates of M.tuberculosis
Microbios 89 135-141 1997 TRC study
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SuggestionsSuggestions
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Role of IQC and EQAP
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Res. Pattern of strain Results obtained
Drug Pattern No. of tests
Agreement
No. %
S
R
S232369
217367
93.599.5
HRS
381299
376286
98.795.7
RRS
384307
382305
99.599.3
E RS
282384
272383
96.599.7
K R S
46169
44168
95.799.4
OflRS
38219
36219
94.7100.0
TOTAL
TRC:IQC IN DST (June’98–Dec 2001)
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Res. Pattern of strain Results obtained
Drug Pattern No. of tests
Agreement
No. %
S
R
S334329
334323
10098.2
HRS
567138
567137
10099.3
RRS
417294
416292
99.899.3
E RS
268458
258457
96.399.8
K R S
28266
27261
96.498.1
OflRS
67296
63292
94.099.8
TOTAL
TRC:IQC IN DST (Jan’2003 – Dec 2004)
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The role of DST in DECThe role of DST in DEC
Failures of category II cases under DOTFailures of category II cases under DOT
Tests should be very simple & rapid for
Primary culture
Identification &
DST
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DST in DECDST in DEC Drug resistance surveillance
The tests should be as per global DRS guidelines
Identification Growth rate. Growth in 500 micrograms of PNB medium Niacin test / NO3 reduction test
DST Indirect economic variant of PST Other methods
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Review of simple & rapid tests DST Review of simple & rapid tests DST for DECfor DEC
DirectDirect Primary culture
Sputum swab method Sputum deposit after processing by Petroff’s
Identification Growth in 500 micrograms of PNB medium
DST Standardization of direct PST only for H & R Absolute concentration method Resistance ratio method
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? RIF. Resistance as an indicator of MDR TB
Direct methods
MABA
Nitrate reductase assay
MTT Assay
MODS
PhaB & Others
Role of speedier pheneotypic methods
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DEFINITION OF RESISTANCE ON LJDEFINITION OF RESISTANCE ON LJSIMPLIFIED VARIANT – PST*SIMPLIFIED VARIANT – PST*
-----------------------------------------------------------DRUGS CONC.(µg/ml) PR (%)------------------------------------------------------------------------ INH 0.2 1 Strep. 4 10 Thioacetazone 2 10 ETH 20 10 Kana 20
10 Cyclo 30 10 Vio 30 10 Capreo 20 10 PZA 100 10 Emb 2 10 Rif 40 1-----------------------------------------------------------------------Only one conc. of drug Canetti et al. Bull. WHO. 1969
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DRS – Salient ObservationsDRS – Salient ObservationsDRS – Salient ObservationsDRS – Salient Observations Among new cases : No evidence of an increase in the
prevalence of resistance
Reports on higher prevalence of ADR ( TRC findings, Gujarat, N.Arcot, N.Delhi, Tamilnadu ,Bombay, UP.)
TRC studies :Low level prevalence of MDR TB
TRC studies : Paediatric & Extra-pulmonary cases• low level resistance to H (5-10%)
• low level resistance to S (2-14%)
• absence of MDR TB
Compared to global situation
• a lesser prevalence of primary resistance
• a much higher level of acquired resistance is
observed
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Issues to be consideredIssues to be considered Steps and Time Tables
Preparation of SOP
Culture system and methodology
Training
Organizing EQAP for second line drugs
IQC Measures (Drugs, Techniques, Periodicity, Monitoring)
Role of Speedier methods for DST of 2nd line drugs
Role of simpler phenotypic methods for detecting MDR TB
Rif. Resistance as an indicator for detecting MDR TB
DST for PZA – Its relevance
Multi - centric approach for defining resistance to various 2nd line drugs by different test systems
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