الفهرس 
TuberculosisOnly 14 pages are availabe for public view
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Abstract
Tuberculosis (TB); infectious disease, which was
considered as a challenging global health problems
especially in developing countries with greater attention
to multidrug-resistant TB (MDR-TB). MDR-TB could be
defined as resistance to isoniazid (INH) and rifampicin
(RIF), two therapeutic drugs in the first-line TB
regimens. (Ahmad and Mokaddas, 2009)
Worldwide, about 3.3% of new TB cases and up to
20% of already treated cases have MDR-TB, and in some
places as Estonia, and Henan Province, China, the
percentage among the new cases increased up to 14 and
11 % respectively. (WHO, 2015) (Dye et al., 2002)
The diagnosis of active TB depended on clinical
suspicion, chest radiographs, and smears for acid fast
bacilli (AFB), then confirmed by solid or liquid culture
which still the “gold standard” reference for TB
diagnosis. However, this method was time consuming
and took several weeks to be completed, so, rapid and
accurate methods for detecting MDR-TB isolates was
mandatory for better controlling and proper treating the
situation from the begging. (Mitchison, 2005) (Sali et
al., 2015) (Mani et al., 2014) In recent years, several molecular methods had
been developed for detection of mutations in rpo B and
kat G genes that caused resistance to INH and RIF,
Cepheid Xpert MTB/RIF kit and the line probe assays
were considered as examples of these methods which
have been commercialized and used in several centers.
(Boehme et al., 2010) (WHO, 2008)
This molecular method gave the physician the
opportunity to detect the resistant bacteria more earlier
and enhance the screening and follow up of these groups
of patients, from this point of view the objectives of the
current work was determined to evaluate the role of
GenoType MTBDRplus assay- as one of commercialized
molecular methods for MDR-TB- in detecting MDR-TB
from sputum of positive TB cases.
The present study conducted on Abbasia Chest
Hospital, TB Department and included 50 newly
discovered positive cases for Mycobacterium
Tuberculosis (diagnosed by direct smear), as a cross
sectional study, the enrolling subjects fulfilled the
inclusive and exclusive criteria of the study; Inclusion criteria:
c) Patients with positive sputum smear for acid-fast
bacilli.
d) Associated clinical and radiological findings
suggestive for pulmonary tuberculosis disease.
Exclusion criteria:
d) Subjects with previous anti-TB failure or relapse.
e) Patients with clinical or radiographic findings
consistent with other chronic respiratory disease.
f) Patients with smear negative even with clinical and
radiological finding suggestive TB infection.
After all subjects had been giving a written
consent prior to participation, the study protocol
approved by the Institutional Research and Medical
Ethics Committees of Ain-Shams University, then all
subjects underwent to the following;
1- Full history taking including; age, sex, smoking
status, addiction habits, co- morbidity
especially DM, Liver and renal disease, past
history of HIV infection or medication used for
it and contact with member diagnosed with
MDR TB.2- Clinical examination.
3- Chest radiography (X-ray)
4- Routine laboratory investigations including;
CBC, Liver function test, Kidney function test
and viral serology for HCV, HBV and HIV.
5- Microbiology assessmentof sputum
including;Ziehl–Neelsen (ZN) staining and
direct microscopy for acid fast bacilli (AFB)
The positive sputum sample for acid-fast bacilli
subjected to:
4- Sputum culture on Lowenstein-Jensen (L-J)
media: (as gold standard test).
5- Drug sensitivity test; (DST) for Rifampicin and
INH by using the proportion method (Heifets,
2000).
6- Mutation detection including;
d) Genomic DNA isolation of the MTB from
sputum samples using QIAGEN QIA amp
DNA mini kit.
e) Genotype analysis by Real-Time Polymerase
Chain Reaction(RT-PCR)strip technology using
GenoType MTBDR plus assay (Hain
Lifescience, Nehren, Germany) Statistical
analysis The data collected, tabulated and statistically
analyzed using IBM SPSS statistics (Statistical
Package for Social Sciences) (V. 22.0) software version
22.0, IBM Corp., Chicago, USA, 2013.
The results summarized in the following points;
1. The mean age of the studied patients was (37.54
±11.06) years, (70 %) of them were males, (54 %)
were smoker, (34 %) were addict to different form
of addiction; the majority were oral tablets (16 %),
then inhalation (10 %) and finally (8 %) were IV
drug addict
2. from the fifty TB patients (22 %) of them gave
history of contact with MDR-TB cases, DM
recorded in (30 %) of cases as commonest
comorbidity presents followed by Liver disease
(18 %), renal disease and malignancy (8 and 4 %)
respectively.
3. Cough, expectoration and toxic symptoms were
the common presentation of patients (98 and 96
%), while haemoptysis presented in (14 %) of
patients.
4. The majority of patients presented by unilateral
lesion in X-ray (66 %) and (30 %) of patients had bilateral lesion, only 2 cases (4 %) had free chest
X-ray.
5. The CBC profile of patients showed; low mean Hb
level (9.30 ± 2.13) mg/dL, with normal mean TLC
and PLT value (9.76 ± 5.16) *103 cell/uL and
(246.60±157.10) *103 cell/uL respectively.Finally
ESR elevated than normal values.
6. HCV test was the commonest positive test (14 %)
of patients, followed by HIV (10 %) and the least
one was HBV (4 %)
7. The prevalence of primary MDR-TB was (12 %)
by molecular methods of detection opposite to (8
%) by conventional (culture) methods.
8. The prevalence of mono drug resistance was (28
%) of patients by molecular methods while by
conventional way was (30 %).
9. Rifampicin resistance alone detected in (8 %) of
patients by molecular methods, while in culture
methods was (10 %)
10. INH resistance alone recoded in (20 %) of
patients by both molecular and conventional
methods
11. The sensitivity, specificity, PPV, NPV and
accuracy of the molecular methods in detection of Rifampicin resistance were (100, 97.5, 90, 100 and
98 %) respectively, and in detecting INH
resistance were (100, 94.4, 87.5, 100 and 96 %)
respectively.
12. Patients characters associated with DR TB
infection showed that;
Male gender was significantly associated with
infection by DR TB, (P = 0.01), with insignificant
statistical difference as regarding the age, (P =
0.8).
No effect of smoking or addiction habits or any
comorbidity on DR development.
Positive contact history with MDR patients had
significant impact on DR TB infection, (P = 0.008)
The patients whom infected with DR TB bacteria
were significantly had lower Hb level than the
other group, (P = 0.009)
Co-infection with HIV, HCV and HBV had
insignificant association with DR TB infection, (P
= 0.3, 0.4 and 0.5) respectively.