RESEARCH PROJECTS

1.         Framework for TB care in Prisons.

A project titled “Framework of TB care in prisons” is being conducted by NDTB Centre. Aim of this project is to estimate the burden of TB among prison inmates at Tihar Jail and to frame the operational modalities for early detection of TB and drug resistant TB at Tihar Jail. Lastly, to mobilize standardized TB care practices among the jail inmates. After sensitization of medical and para medical staff of Tihar, Active Case finding was started from the month of October 2018.   Till 31st March,2019. 10476 inmates have been screened, out of which 655 symptomatics were found and 70 TB cases were detected.  All detected TB cases have been put on treatment as per RNTCP guidelines.

 

2.         Active Case Finding Campaign of Tuberculosis among People who use           intravenous Drugs in New Delhi.

Active case finding campaign was an activity propagated by the programme. This was a house to house survey for search of TB symptomatic .The Objective of the OR is (a) Early identification of TB suspects. (b) Early diagnosis (c)Treatment preparedness (d) Link to Treatment and Care. Targeted Population : Person who Inject drugs in the area of Delhi of Yamuna Bazar and Hanuman Mandir in CP of New Delhi. Estimated number of population will be from field area of two Target Intervention centres of DSACS project which is approximately 2500. After training of the field teams, Active Case finding will be started. Recruitment of field workers, their sensitization and micro planning has been done.

 

3.         TB free Rohini Project

Keeping in mind the target of TB elimination by 2025, a project has been initiated to make Rohini TB free.  The project is jointly conducted by IMA Rohini Branch, NDTB Centre and Delhi State TB Department.  It started with sensitization of local politicians, Private practitioner and community of Rohini area in the month of March 2019.  Further activities have been planned to conduct active case finding in the area, increasing TB notification by private practitioners and utilizing local politicians and community for treatment adherence of detected TB cases.

 

4.       Drug susceptibility testing of Mycobacterium tuberculosis isolates against Isoniazid and Rifampicin using Line probe Assay

          A study was carried out from February 2019 to April 2019 at the Intermediate reference laboratory of New Delhi Tuberculosis Centre, New Delhi, with following objectives:

 

a) Isolation of Mycobacterium tuberculosis from smear negative pulmonary specimens using MGIT liquid culture system. b) Detection of drug resistance to first line anti TB drugs isoniazid and rifampicin using molecular line probe assay.

          In this study,  615 sputum samples from Presumptive MDR TB patients belonging to 17 out of 25 Chest Clinics of Delhi were  collected in sterile, and leak proof containers  and were  transported  on same day to IRL  along with  filled requisition form. These samples were decontaminated using NALC-NaOH method. From concentrated sediment, smears were prepared and were stained using Auramine stain. By using Fluorescent microscopy, smears were read as negative and positive. Smear negative samples were further subjected to culture on MGIT 960 system. Confirmed positive cultures were further processed for LPA to detect resistance to first line drugs INH and RIF.

 

Because of the low sensitivity of sputum microscopy, TB diagnosis is usually confirmed by culture, followed by the identification of the strain and drug susceptibility testing (DST). In this study, LPA was done instead of phenotypic DST which gave results earlier.

 

 

 

 

5.       Isolation, Identification and Drugs Susceptibility testing of Mycobactererium tuberculosis from Extra Pulmonary clinical Specimens

 

          The laboratory diagnosis of EPTB has always been a challenge. Therefore this study was carried out during the period from 1 February 2019 to 1 May 2019 and samples were collected from presumptive extra pulmonary patients belonging to 17 out of 25 Chest Clinics of Delhi.  The objectives of the study were

(A) Isolation of M. tuberculosis from extra pulmonary specimen from presumptive TB cases.

(B) Identification of M. tuberculosis using commercial immuno chromatographic assay.

(C) Detection of drug resistance of first line drugs namely Isoniazid and Rifampicin using molecular LPA.

          Specimens were collected and were transported, on the same day of collection to the tuberculosis laboratory along with filled RNTCP request form.  These specimens were decontaminated by N-acetyl L- Cysteine Sodium Hydroxide (NALC –NaOH) method and from the concentrated sediments, smears were prepared. The smears were stained with Auramine stain and were read using LED florescent microscopy.  From all concentrated sediments, a portion was inoculated on MGIT medium vials, and these vials were incubated in MGIT system. All culture positive samples were   then identified as M.tuberculosis complex using the immuno chromatographic assay. After identification,   the cultures were used to extract DNA and   these DNA samples were further processed for  first line LPA.

The availability of valid LPA results proved  the advantages of molecular test over the conventional method The assay gave interpretable results and detected an additional  MDR-TB case

6.       Detection of drug resistance for second line anti TB drugs among MDR TB OR Rifampicin resistance case using MGIT liquid culture and drug susceptibility testing

 

         For a better management of drug-resistant cases, early detection of resistance is extremely important so that effective treatment can be prescribed. Rapid drug susceptibility testing plays an important role in the detection and control of MDR/XDR TB. The present study was carried out with the following objectives,

 

a) Detection of drug resistance to rifampicin using commercial Xpert MTB/Rif assay among Pulmonary TB suspects

b) Isolation of Mycobacterium tuberculosis from pulmonary specimens using MGIT liquid culture system

c) Detection of drug resistance for second line drugs using MGIT liquid culture system.

 

         Fresh sputum Samples were collected in sterile and leak proof containers at DMC and transportation was done on same day to IRL along with patient information form.  These samples were decontaminated by NALC-NaOH Method. Sediments were used to prepare smears and also for culture inoculation. Fluorescent Microscopy was done using Auramine stain and the smear were read as per RNTCP guidelines.

 

The BACTEC 960 TB System was used for cultivation of M. tuberculosis.  For all positive cultures, Identification of M. tuberculosis complex was done using MPT64 rapid test. Second line Drug Susceptibility Testing using MGIT 960 system was done using six drugs  Kanamycin, Amikacin, Capreomycin, Moxifloxacin, Linezolid and Clofazimine.

 

          This study results showed that liquid culture offers a more sensitive and rapid method for isolation of M. tuberculosis and performing susceptibility testing against a variety of first-line and second-line anti tuberculosis drugs

 

7.       Detection of drug resistance to second line anti TB drugs among Rifampicin resistance TB cases using molecular Line Probe Assay

         The present study was carried out with following objectives

a)                       Detection of drug resistance to rifampicin using commercial Xpert MTB Rif  assay among pulmonary presumptive TB cases.

b)                        Detection of drug resistance to second line injectables and fluoroquinolones  using  Molecular Line Probe Assay.

.        Fresh sputum Samples were collected in sterile and leak proof containers at DMC  from  patients diagnosed as Rifampicin resistance by CBNAAT. Transportation of samples  to IRL was done on same day to IRL with patient information form.

NALC-NaOH Method: These samples were decontaminated by NALC-NaOH Method. Sediments were used to prepare smears and also for culture inoculation

Microscopy:  Smears were prepared using Grease free slides,. Staining was done using Auramine. Smears were read by using LED flurescence microscopy as per RNTCP guidelines

Line probe assay;   Line probe assay was performed for second line drugs using GenoType MTBDRsl VER 2.0 LPA.

It was observed in this study that   second line LPA performed well with excellent reproducibility and high sensitivity and specificity for FLQ and SLID resistance determination. The average reporting turnaround time varied from 2 to 4 days.