S. Vashakidze, N. Shubladze, L.Kupreishvili,K.Nikolaishvili, L. Mskhiladze, I.Khurtsilava,

*J. E. Graham, 

National Center of TB and Lung Diseases, Tbilisi,Georgia

And *University of Louisville School of Medicine, Louisville, KY

University of Louisville Jim Graham

Gene arrays Sorting the specimens

MiniCycler RT-PCR machine

Explaining the poster Almost ready

Goal:Comparative analysis of drug-resistance in tubercle bacilli recovered from both patient sputa andlung tissue resectates at the Georgian National Center of TB and Lung Diseases.Materials and methods:  M. tuberculosis isolates were obtained from a total of 27 patients undergoingsurgical intervention therapy in the Surgery Department of the Georgian National Center of TB andLung Diseases. All patients had been treated with previously unsuccessful conventional drug therapiesfor from 6 months to several years prior to surgery.Standard sputum microscopy and cultural analyses were performed along with first line (isoniazid andrifampin) susceptibility testing both before and after surgery. Lung tissue resectates were taken duringsurgery and designated as either ‘I’ for internal wall of cavitating granulomas, ‘E’ for external wall (orpericavital area), ‘C’ for caseous mass (necrotic material inside cavity), or ‘N’ smaller peripheralnodulus often seen in distinct lung tissues. Studies were supported by U.S. NSF-CRDF grant #GEX1-002712-TB-06, and carried out as approved by local IRBs.Results: Out of 27 patients before operation, 11 (40.8%) were sputum-smear positive, and 16 (59.2%)were sputum-smear negative. M. tuberculosis was successfully cultured from sputa for 19 patients (70.3%). Tubercle bacilli were also cultured from lung tissues obtained during surgery in 14 cases (51.8%).The frequency of successful culture differed in different types of pulmonary lesions and areas withinlesions, with 92.8% culture positive from C, 78.5% for I, 71.4% for E, and 41.6% for N. Comparative analysis of DST-s from sputum and surgical materials showed following results:

In sputum : 1 isolate was pansensitive, monoresistance in 0 cases, polyresistance in 7, multiresistance in 5, total resistance in 1 isolate.

In resectates: monoresistance-1 isolate, polyresistance-3, multiresistance-13, total resistance-9.Conclusions: Isolates obtained from lung resectates appeared to have higher resistance profile than obtained

from the sputum of the same patient. It should be considered in creating of treatment strategy in post-surgery period.

Operation specimens from 27 patients : ‘I’ for internal wall of cavitary granuloma ‘E’ for external wall ‘C’ for caseous mass (necrotic material

inside cavity) ‘N’ smaller peripheral

nodulus

Cavity Tuberculoma

Ziel-Nielsen microscopy

Drug Susceptibility testing by proportional method on Lowenstein-Jensen medium

Microbiology data

Sputum specimens

Before operation After operation

AFB+ AFB- AFB+ AFB-

Microscopy 11 (40,8%) 16 (59,2%) 0 27 (100%)

Culture 19 (70,3%) 8 (29,7%) 0 27 (100%)

All bacterial cultures were identified as M.tuberculosis.Before operation:

0

20

40

60

80

Microscopy Culture

AFB+

AFB-

AFB+

AFB-

Specimen I C E N

Frequency 11/14 13/14 10/14 5/12

78,5% 92,8% 71,4% 41,6%

Quantity 2,1+ 1,8+ 1,5+ 1,5+

0

2

4

6

8

10

12

14

Monoresistant

MDR Poly resistant Totallyresistant

Sputum

Resectate

Specimen site Mono resistant

MDR Polyresistant

Totallyresistant

Sputum 0 5 10 1

Resectate 1 13 3 9

0

20

40

60

80

100

I C E N

I

C

E

N

M. tuberculosis capable of forming colonies on L-J slants were most readily detected in necrotic caseous material, to a lesser extent in both inner and outer cavity walls. Less frequently, live bacteria were found in small nodules. Further investigation will allow comparing patterns of M. tuberculosis gene expression during growth in the human lung.

Isolates obtained from lung resectates appeared to have higher resistance profile than obtained from the sputum of the same patient. It should be considered in creating of treatment strategy in post-surgery period.

In case of MDR- M.tb finding in operation specimens it is necessary to prolong treatment with antituberculous drugs for at least 18 months after the operation.

NCTLD UofL