Journal of Biomedical and Biological Sciences

 

Haematological and Biochemical Profile, Risk Factors Analysis, and Sensitivity Comparison between Microscopy and Gene Xpert MTB/RIF Assay Method for Identification of Mycobacterium tuberculosis

Rida Maqbool1, Abdul Wajid1, Muhammad Ishfaq2*ORCID ID and Shaheen Qasim3 

1Department of Biotechnology, Faculty of Science and Technology, Virtual University of Pakistan, Lahore, Pakistan.

2Institute of Molecular Biology and Biotechnology, Bahauddin Zakariya University, Multan, Pakistan. 3Department of Biochemistry & Biotechnology (Baghdad-ul-Jadeed Campus), The Islamia University of Bahawalpur, Pakistan.

*Corresponding Author: Ishfaq M, Institute of Molecular Biology and Biotechnology, Bahauddin Zakariya University, Multan, Pakistan. E-mail: mi742813@gmail.com

Citation: Maqbool R, Wajid A, Ishfaq M and Qasim S. Haematological and Biochemical Profile, Risk Factors Analysis, and Sensitivity Comparison between Microscopy and Gene Xpert MTB/RIF Assay Method for Identification of Mycobacterium tuberculosis. Journal of Biomedical and Biological Sciences. 2021;1(1):1-14.

 

Copyright: © 2021 Ishfaq M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Received On: 5th November,2021    Accepted On: 14th November,2021    Published On: 24th November,2021

Abstract

Mycobacterium tuberculosis is a major health problem because of its diverse treatment plan and management for health of public for developing countries like Pakistan and worldwide. Approximately 1 in 16 patients is affected from Mycobacterium tuberculosis. Transmission of Mycobacterium tuberculosis is through air droplets containing the infectious bacteria and so much lethal bacteria that even a single microorganism is sufficient to infect the healthy person. But if it is diagnosed timely at initial stages of disease through molecular base new technology GeneXpert assay then it can be cured easily with Anti-tuberculosis drugs and also its transmission can be controlled. Zheil Nelson (ZN) microscopy method and chest X-ray are two diagnostic tests commonly used for Mycobacterium tuberculosis finding. The GeneXpert assay was developed for determination of Mycobacterium tuberculosis and drug resistance TB. In the developing countries, conventional methods such as sputum microscopy is mostly used instead of GeneXpert assay, thereby yielding false positive and false negative results as well as accomplish comparatively long time in diagnosis of Mycobacterium tuberculosis. A cross sectional study on 1000 patients was the part of this study to check epidemiology, haematological and biochemical spectrum and the risks associated with pulmonic tuberculosis & persons affected with MDR Tuberculosis as well as to compare the sensitivity of Fluorescent, ZN microscopy and GeneXpert method for identification of Mycobacterium tuberculosis. There was total 682 included in this study. Majority of them from poor socio-economic status (84%) and due to this factor, they have also poor nutritional status. These patients were living in areas where access to healthcare facility was not easy and 70% of them had positive smoking history. There were 682 patients reported with 171 new cases and 643 were with MTB at district Bahawalnagar. There was total 15 patients which were retreated while 39 patients were from follow-up. The highest sensitivity was of Gene Xpert, while specificity of all three tests Fluorescent Microscopy, ZN Microscopy and Gene Xpert were equal (99). Positive Predictive Value of Gene Xpert and Fluorescent Microscopy were equal (98) and high from from ZN Microscopy.

Keywords: Mycobacterium tuberculosis, GeneXpert assay, ZN microscopy, Chest X-ray, epidemiology.

Introduction

Tuberculosis is the major contiguous infections all over the world and it is caused by Mycobacterium tuberculosis bacterium that has effects on lungs [1]. Tuberculosis is supposed to be the second leading reason of deaths from transmitted contagious diseases after the human immunodeficiency virus. This is one of the oldest diseases that occurred with humanity during its development. Many studies in Europe and the Middle East have been found which demonstrate the presence of the disease’s etiologic agent for 4,000 years [2]. In 2013, World tuberculosis report showed the total 8.6 million cases and 1.3 million deaths. This data indicated that, in spite of many efforts made by government organizations to combat with this disease, it continues to be a serious community health issue over the worldwide [3]. In current years, many of the advance molecular biology techniques have led to changes in the genetic codes of several microorganisms, such as M. tuberculosis to facilitate the investigation of tuberculosis, plug-in elements use polymorphism and analysis in the relapse control line Direct (DR) with analysis. Repeating units of intersection of mycobacterium (IS6110) allows to vary the level of knowledge about tuberculosis, which contributes to the growth of epidemiology (molecular) and disease control policies in different cities and countries [4].

The World Health Organization (WHO) estimates that about 1/3 of the world’s population is M. tuberculosis [5]. In 2000, 8 to 9 million cases of the disease and about 3 million cases of tuberculosis died worldwide. After the human immunodeficiency virus (HIV) / AIDS, tuberculosis is the second most common cause of death from an infectious disease, and current trends prove that this year will be among the top 10 causes of tuberculosis worldwide [6].

The global distribution of tuberculosis is strongly biased towards emerging and low-income countries. Asia has the largest cassia in Asia, with China, Indonesia, Bangladesh, India and Pakistan together accounting for more than 50% of the world’s weight. Africa and especially sub-Saharan Africa have the highest number of TB cases per 60,000 and 100,000 per 100,000, respectively. Tuberculosis usually affects the age group of 15-25 years (about 6 million out of 8 million) in the most economically productive age group. Our understanding of tuberculosis epidemics and the effectiveness of control mechanisms is complicated by the emergence of drug-resistant bacilli and the association of tuberculosis with HIV co-infection [7].

The emergence of tuberculosis isolates has been observed. In vitro studies have shown that m. Spontaneous changes in tuberculosis may be associated with drug resistance, while selective (antibiotic) stress may result in further accumulation of these drug-resistant mutants. The effective choice of drug resistance in the presence of a single antibiotic prompted researchers to recommend combination therapy with multiple antibiotics to reduce the incidence of drug resistance during treatment. In fact, when there is an adequate supply of drugs and combined treatment is properly organized, the fight against tuberculosis is effective [8].

The selection of drug-resistant mutants in patients occurs primarily when patients receive inadequate treatment or are temporarily exposed to sub therapeutic levels of the drug, a situation that may arise and provide adequate positive selection pressure to maintain. De Novo is a drug resistant organism. A remarkable time of chemotherapy is required for the treatment and cure of tuberculosis infection.

The need for the agent to maintain high levels of the drug for several months of combined treatment, including innate toxicity, reduces patient adherence and therefore increases the likelihood of drug resistance. Therefore, the need to identify new anti-TB agents as well as reduce the duration of chemotherapy is universal, as it will have a major impact on clinical management and the emergence of drug resistance. Since the early 1990s, one of the main sources of dangerous trends and public health problems has been the emergence of multi drug resistance (MDRTB), two of the most resistant, defined as isolated (INH) and rifampicin (RIF).

Recent estimates show that there were 458,000 cases of MDR-TB worldwide in 2003 (including new and re-treatment cases) (951 confidence breaks, 321,000 to 689,000). These data show that incessant events can be two to three times more common than events and that a very large number of people have recently been infected. Although MDR-TB treatment has improved significantly (primarily in resource-rich settings), treatment is usually more difficult, with much higher morbidity and mortality associated with long-term treatment and increased risk of recurrence. It happens. Resistant to community isolated drugs [9].

Objectives of the present study was to gather Haematological and Biochemical profiles variations analysis in pulmonary tuberculosis patients and MDR TB patients’ samples, to find out the sensitivity of Gene Xpert MTB/RIF assay for Mycobacterium tuberculosis bacteria recognition in sputum samples. The objective also includes a comparison of Gene Xpert MTB/RIF assay with Fluorescent and ZN Microscopy Method technique for the analysis of Mycobacterium tuberculosis detection.

Materials and Methods

Methodology/Research Design

A cross sectional study was carried out at DHQ Hospital Bahawalnagar in southern Punjab, Pakistan.

During research (From July 2019 to November 2019), 1000 sputum samples were taken from Mycobacterium tuberculosis patients. Total 300 Blood samples were collected from MTB (Mycobacterium tuberculosis) patients to study the Haematology and Biochemistry parameters changes.

Criteria for sample collection

Presumptive TB case /patients suspect clinically having TB having (cough of 2 weeks, weight loss, fever or chest x-ray having image of TB) Sputum samples of persons suspecting for TB, advised by the relevant doctor of TB clinic.

Sampling protocol

Sputum and blood Samples were collected as per standard protocol of sputum/Blood collection in TB lab. Blood samples were collected in EDTA vials. Samples of patient were taken after proper identification, labelling, name, age, MR number or brief medical history. Data of the patients related to age, gender, socio-economic status, life style and education from all patients was collected through questionnaire.

Pulmonary sample

The sputum samples from patients were collected early in the morning as per standard protocol. There are two types of sterilized sputum container given to the TB patient for the collection of sputum at their homes or hospital. Ziehl Neelsen staining and Auramine Staining (Fluorescent Microscopy) was performed as per standard methods.

Blood sampling

Blood samples from TB patients were taken as per research requirement for Haematology and Biochemistry profiles in TB patients. A sterile syringe was used aseptically to draw peripheral blood almost 5-6ml. CBC vial containing 0.2 ml of 4% EDTA to get 2ml of blood to check different parameters (total leucocytes count, RBCs count, Platelets count and peripheral smear for differential Leucocyte’s count) through haematology analyser (Sysmex, KX-21). Approximately 2 ml for Erythrocyte sedimentation rate (ESR) determination blood having anticoagulant was tested. Readings were taken from tube standing vertically into stand. After that remaining 1-2 ml clotted blood sample were used for analysis in the Biochemistry Analyzer (Micro Lab 300) for evaluation of different biochemical parameters (liver enzymes).

Gene Xpert Assay test

This was performed directly on sputum pellets or raw sputum samples, samples after decontaminating and concentrating the sputum were considered best sample for test accuracy. Clinical sputum samples are mixed with a sodium hydroxide and isopropanol. Then this treated sputum samples are added in a ratio of 3:1 for sputum pellets and incubated at room temperature for duration of 15 min. This step is intended to decrease the viability of M. tuberculosis in sputum and to decrease biohazard. The treated sample is then manually transferred to the cartridge machine which is then transferred into the PCR base GeneXpert instrument.

In Gene Xpert test measured amount of 2ml of sputum sample was added to the sample well in given cartridge. The cartridge with added sample is placed in the instrument then programmed ultrasonic lysis of the bacilli and release of the genetic material.  The cartridge encountered with a syringe drive, a rotary drive and further procedure of amplification started internally in the machine. After 2 hour of this procedure the detection of MTB displayed on screen as detected or not detected MTB.

The Gene Xpert assay amplified segment of the Rifampicin resistance region of Mycobacterium tuberculosis detected using real time-PCR reaction and the assay also contains lyophilized Bacillus globigii spores which serve as an internal sample processing and PCR control (SPS). Molecular beacons /Florescent probes which Complementary to the Rifampicin resistance determining region named as Primary probe D, probe C, probe E, probe B and probe A used. By fluorescent probes, gene mutation anywhere present in the core region of RRDR for RIF resistance results in either partial inhibition or complete suppression of corresponding molecular beacon and RIF Resistance detection were seen as in result in this assay detected or not detected or indeterminant resistance and which gene is involved.

There are various generations of Gene Xpert assay such UltraXpert now has revolutionized. Sputum sample is mixed with diluents chemical and after incubation of 20 min at room temperature. Diluted sputum sample of 2ml is added into the cartridge. Then this cartridge was loaded in the instrument. Kits and reagents used for Gene Xpert assay should be stored at 2-8oC.Because the sensitivity and reliability of test results also depend on the storage and carefully handling of kits, chemicals and reagents used during test.

Statistical Analysis

Comparison of two techniques was elaborated by ANOVA tables. Analysis of clinical parameters results and demographic characteristics are presented as frequency percentages and mean + S.D. based on their categorical and numeric nature, respectively. Frequency percentages, tables and graphs using appropriate statistical analysis done by statistical tool SPSS20.

Ethical Approval

Consent of patients and attendant was taken to allowing us to use their sample for research purpose.

Results

This research work was aimed to evaluate the molecular epidemiology, haematological, biochemistry profile and risk factors analysis and sensitivity comparison between microscopy and gene xpert MTB/RIF assay method for identification of pathogenic Mycobacterium tuberculosis in district Bahawalnagar.

Socio-demographic profile of the patients of Tehsil Bahawalnagar

There were total of 682 patients enrolled from tehsil Bahawalnagar, out of 682, majority (472) were male patients and 210 were females.

 

Figure 1: Gender Distribution of patients collected in this study.

There was total 682 included in this study. Majority of them from poor socio-economic status (84%) and due to this factor, they have also poor nutritional status (Figure 2). These patients were living in areas where access to healthcare facility was not easy and 70% of them had positive smoking history. While chronic pulmonary diseases were the main factors associated in patients.

 

Figure 2: Risk Factors analysis

There were 682 patients reported with 171 new cases and 643 were with MTB at district Bahawalnagar. There was total 15 patients which were retreated while 39 patients were from follow-up.

 

Month

Total cases

MTB +ve/ New patient

Retreated/follow-up

RR

MTB negative

July

129

41/119

05/10

0

73

Aug

95

28/89

2/6

1

53

Sep

106

22/99

3/7

1

78

Oct

188

47/178

2/10

1

136

Nov

164

33/158

3/6

2

120

Total

682

171/643

15/39

5

460

Table 1: Month-wise sample collection from Tehsil Bahawalnagar

 

Figure 3: Month-wise patients reported at tehsil Bahawalnagar

Month wise patient’s data recorded. Highest number of patients were in the month of October (188), followed by November (164).

 

Figure 4: Month-wise MTB positive cases at tehsil Bahawalnagar

Month wise MTB positive cases were assessed, and highest number of MTB positive cases were reported at the month of October (49) while lowest number of cases were reported in the month of July (46) at district Bahawalnagar.

 

Tuberculosis

ZN

FM

Gene Xpert

MTB Positive Cases

164

201

278

MTB Negative Cases

10

16

13

Table 2: Comparison between diagnostic methods for MTB Positive and negative cases distribution at Tehsil Bhawalnagar (n=682)

There were 164 MTB positive cases on ZN, 201 on FM and highest value was on Gene Xpert (278). While highest negative cases were on FM (16).

 

 

 

Percentage

 

 

Positive

Negative

Sensitivity

Specificity

PPV

NPV

Agreement

ZN Microscopy

Positive

94

4

39

99

96

63

14.5

Negative

255

329

Fluorescent Microscopy

Positive

171

2

55

99

98

72

79

Negative

137

372

Gene Xpert

Positive

306

5

98

99

98

99

98

Negative

4

367

Table 3: Sensitivity, specificity, PPV and NPV of different diagnostic modalities at Tehsil Bhawalnagar (n=682)

There were calculated sensitivity, specificity, positive predictive value and negative predictive value. The highest sensitivity was of Gene Xpert, while specificity of all three tests ZN Microscopy, Fluorescent Microscopy and Gene Xpert were equal (99). Positive Predictive Value of Gene Xpert and Fluorescent Microscopy was equal (98) and high from ZN Microscopy.

Socio-demographic profile of the patients of Tehsil Chishtian

There was total 318 included in this study from Tehsil Chishtian. Majority of them from poor socio-economic status (78%) and due to this factor, they have also poor nutritional status. These patients were living in areas where access to healthcare facility was not easy and 78% of them had positive smoking history having chronic pulmonary diseases main factors associated in patients.

 

Figure 5: Gender distribution of patients

There was total 318 patients enrolled in our study. Out of 318, majority of patients were male (202).

 

Month

Total test

New patients

Follow-up/retreated

MTB +ve

Rifampicin Resistance cases

MTB -ve

July

19

19

0

14

01

5

August

51

51

0

40

02

11

September

68

68

0

51

01

17

October

109

109

0

88

02

21

November

71

71

0

56

01

15

Total

318

318

0

249

07

69

Table 4: THQ Chishtian New and MTB positive cases

There was total 318 patients came at THQ Hospital Chishtian. Out of 318, there were 249 patients were MTB positive cases, while 69 were MTB negative cases. There was not patient from follow up or retreated category.

 

Figure 6: Month-wise patients reported at Tehsil Chishtian

Month wise reported cases were assessed, and highest number of MTB positive cases were reported at the month of October (88) while lowest number of cases were reported in the month of November (56) at Tehsil Chishtian.

 

Figure 7: Month-wise MTB positive cases at Tehsil Chishtian

Month wise reported cases were assessed, and highest number of MTB positive cases were reported at the month of October (88) while lowest number of cases were reported in the month of July (14) at Tehsil Chishtian.

 

Tuberculosis

ZN

FM

Gene Xpert

MTB Positive Cases

57

89

113

MTB Negative Cases

13

21

25

Table 5: Comparison between diagnostic methods for MTB Positive and negatives cases distribution at Tehsil Chishtian (n=318)

MTB cases were diagnosed through ZN, FM and Gene Xpert. Highest number of cases were diagnosed through Gene Xpert (113), followed by FM (89) and lowest were from ZN (67).

 

 

 

Percentage

 

Positive

Negative

Sensitivity

Specificity

PPV

NPV

Agreement

ZN Microscopy

Positive

43

2

39

99

96

63

14.5

Negative

134

139

Fluorescent Microscopy

Positive

80

3

55

99

98

72

79

Negative

70

165

Gene Xpert

Positive

162

5

98

99

98

99

98

Negative

2

149

Table 6: Sensitivity, specificity, PPV and NPV of different diagnostic modalities at Tehsil Chishtian (n=318)

There were calculated sensitivity, specificity, positive predictive value and negative predictive value. The highest sensitivity was of Gene Xpert, while specificity of all three tests ZN Microscopy, Fluorescent Microscopy and Gene Xpert were equal (99). Positive Predictive Value of Gene Xpert and Fluorescent Microscopy was equal (98) and high from ZN Microscopy.

 

 

Sum of Squares

Df

Mean Square

F

Sig.

Between Groups

318

3

156.4

8.3

0.00

Within Groups

682

28

67.83

 

Table 7: ANOVA Results

There was found F with value of 8.3 with having 0.000 p-value which showed the results significant. Different diagnostic techniques found difference of means statistically significant.

 

Month

Bahawalnagar

Chishtian

MTB +ve

MTB +ve

July

46

14

August

30

40

September

25

51

October

49

88

November

36

56

Total

186

249

Table 8: Total MTB positive cases at Tehsil Bahawalnagar and Chishtian

 

Month

Total test

New patients

Follow-up/retreated

MTB +ve

Rifampicin Resistance cases

MTB -ve

July

148

148

05/10

60

01

78

August

146

146

2/6

70

03

64

September

174

174

3/7

76

02

95

October

297

297

2/10

137

03

157

November

235

235

3/6

92

03

135

Total

1000

1000

15/39

435

12

529

Table 9: Total Rifampicin resistance & MTB Cases at Tehsil Bahawalnagar and Chishtian

 

Total Cases/samples

Extra-Pulmonary Cases

21

Pulmonary Cases

979

Table 10: Pulmonary and Extra-Pulmonary Cases at District Bhawalnagar

Reported cases were divided into pulmonary and extra-pulmonary cases. There were very few extra-pulmonary cases (21) while remaining cases were pulmonary (979).

Laboratory Investigations

Haematology Parameter

Erythrocyte’s sedimentation rate value was recognized under three different categories having (ESR values with 15-50, 51-100 and >100mm/hr). More than 60% of patients belong to the category of ESR 35-50 mm/hr, while 15 cases exceeded the value more than 100 mm/hr.  It was observed that haemoglobin was critically lower than normal values (10-13 g/dl) with 65% in males and 35% in female patients respectively.

The white blood cell counts show some features in a way that total leukocytes count was found to be lower than normal in 10 patients. While lymphocytopenia was found in 166 males and 134 in female patients respectively. Neutrophilia was found 140 in males and 160 in females.

Platelet’s count was found higher in most of the patients; however, it was observed that thrombocytopenia was found in 15% of the patients.

Biochemistry parameter

Bilirubin total among positive cases was slightly raised almost half of the patients. SGPT (ALT) values of positive cases were raised among 135 male and 110 female patients while, SGOT (AST) was raised among 160 males and 148 females.

 

LABORATORY INVESTIGATIONS

HEMATOLOGY REPORTS

Mean ± SD

Hb (g/dl)

10.61 ± 0.94

ESR (mm/h)

75.85 ± 40.4

TC (×103/µl)

7325 ± 1888.53

RBC (×106/µl)

3.49 ± 0.33

Platelet (×103/µl)

1.88 ± 0.34

BIO-CHEMISTRY REPORTS

SGPT (ALT)

Male

135 (13.5%)

Female

110 (10.1%)

SGOT (AST)

Male

160 (16%)

Female

148 (14.8%)

Table 11: Laboratory Investigations

Discussion

The Xpert MTB/RIF assay have 100% sensitivity is considered and compared with gold standard. These observations are same with already done studies [10-12] evaluating the sensitivity of the Xpert MTB/RIF test for the analysis of pulmonary MTB with positive results of smear microscopy. In Earlier studies it has been shown that approx. 71% of smear-negative smears are sensitive to Xpert MTB/RIF assay, which is suggestively less than the sensitivity detected in our study, especially in samples without breathing. This might be because of the sample quality or changes in patient demographics. However, the possibility of this analysis to detect smear-negative MTB is clinically very important, as more and more cases of smear-negative smear are found in tuberculosis patients diagnosed with human immunodeficiency virus (HIV), especially at laboratories with inadequate resources [12]. The Xpert MTB/RIF is also supposed to be more cost effective than all other utilized conventional methods. The countries having high MDR-TB cases, the price of using this Xpert MTB/RIF for diagnostic is 4% of already mention TB funding. In comparison to other high TB in Africa, the actual price of using to diagnose TB in patients have HIV-positive including the TB signs denotes only 1–3% of the fund.

Although the culture technique is counted to be the perfect standard for the analysis of lung MTB infection using a smear-positive test, we found that the Xpert MTB/RIF analysis significantly increases the detection of MTB with greater sensitivity in patients with smear-positive as compared to smear-negative [13]. Which is constant with a meta-analysis of analysis of the effectiveness of Xpert MTB/RIF in multicounty research revealed a sensitivity of 83.1% between culture-positive cases of TB and 98.01% in patients with a positive smear and negative MTB result. Cases of negative smear tuberculosis are associated with delayed or inability to make an analysis, outcomes of poor dealing, danger of transmission and death. The introduction of Xpert MTB/RIF analysis in the initial stages of an analytical workflow can possibly progress detection and shorten laboratory response times. Current reports propose that the Xpert MTB/RIF assay raises the incidence of RIF resistance and reduces needless empirical dealing amongst smear-negative pulmonary MTB smears [14].

Diagnosis of extra pulmonary tuberculosis infection is a serious problem, probably due to the fewer organisms that could be restored in the sample. We found in our study that the Xpert MTB/RIF assay works well in identifying extra pulmonary tuberculosis from different places (lymph node biopsy, peritoneal fluid, pus, and gastric aspirate). Incidence of MTB amongst smear-negative respiratory specimens using the Xpert MTB/RIF test varies between studies with a sensitivity level of 20.00-47.7% [11, 12]. As mentioned above, Xpert MTB/RIF has greater sensitivity for analysing MTB in the lymph nodes (83.00%) as compared to other samples. It may be due to the heavy load on mycobacteria during disease, also the comfort of access to the sampling site. This data is significant in the diagnosis of extra pulmonary tuberculosis, especially in the paediatric age group in which up to 60.00% of patients with extra pulmonary tuberculosis acquire cervical lymphadenitis [15]. In addition, since the identification of MTB in children is every so often delayed because of unspecific clinical characteristics and difficulty in coughing, the Xpert MTB / RIF analysis is a useful method for diagnosing MTB in children.

Although neither the FDA and European Compliance (CE), has agreed Xpert MTB/RIF for the analysis of extra pulmonary MTB, the World Health Organization (WHO) has issued a policy statement mentioning Xpert MTB/RIF for the analysis of extra pulmonary resistance of MTB and RIF in children. This verdict was based at new data from a analysis of 85 reviewed publications that showed that Xpert MTB/RIF works well in perceiving extra pulmonary infections, including lymph nodes, CSF, and body fluids. These references are constant with one more meta-analysis in which 27 reviewed journals collectively propose that Xpert MTB/RIF precisely detects additional pulmonary tuberculosis in children and adults [16].

The Xpert MTB/RIF study stated a false positive in this research. The Patients which showed false-positive results may acquire anti-TB treatment that can be avoided. False positive results were previously reported in the Xpert MTB/RIF study due to the existence of dead MTB in examination samples, particularly amongst formerly treated patients [17-20]. Because of the lack of access to patient records, it is not possible to obtain information about previous treatment with anti-TB drugs. A thorough medical history, with particular emphasis on pre-treatment with anti-TB drugs, is crucial to avoid false positives.

Conclusion

Numbers of MTB cases are not reducing in remote areas of Punjab like District Bahawalnagar and Tehsil Chishtian. There is need of more focused efforts in this regard. This research also verified good sensitivity and specificity of Xpert assay in detecting M. tuberculosis. The treatment of TB is significantly made easy and fast because of this diagnostic test, mostly in those patients who had a negative sputum acid-fast smear.

References

  1. Singh N, Paterson DL. Mycobacterium tuberculosis infection in solid-organ transplant recipients: impact and implications for management. Clinical infectious diseases. 1998 Nov 1;27(5):1266-77. 
  2. Jordao L, Vieira OV. Tuberculosis: new aspects of an old disease. International journal of cell biology. 2011 Jun 21;2011. 
  3. World Health Organization. Global tuberculosis report 2013. World Health Organization; 2013.
  4. Reis AC, Tenreiro R, Albuquerque T, Botelho A, Cunha MV. Long-term molecular surveillance provides clues on a cattle origin for Mycobacterium bovis in Portugal. Scientific reports. 2020 Nov 30;10(1):1-8. 
  5. World Health Organization, Stop TB Initiative (World Health Organization). Treatment of tuberculosis: guidelines. World Health Organization; 2010. 
  6. Zozio T, Allix C, Gunal S, Saribas Z, Alp A, Durmaz R, Fauville-Dufaux M, Rastogi N, Sola C. Genotyping of Mycobacterium tuberculosis clinical isolates in two cities of Turkey: description of a new family of genotypes that is phylogeographically specific for Asia Minor. BMC microbiology. 2005 Dec;5(1):1-9.
  7. Mathema B, Kurepina NE, Bifani PJ, Kreiswirth BN. Molecular epidemiology of tuberculosis: current insights. Clinical microbiology reviews. 2006 Oct;19(4):658-85.
  8. Dai J, Chen Y, Lauzardo M. Web-accessible database of hsp65 sequences from Mycobacterium reference strains. Journal of clinical microbiology. 2011 Jun;49(6):2296-303.
  9. Cavanagh R, Begon M, Bennett M, Ergon T, Graham IM, De Haas PE, Hart CA, Koedam M, Kremer K, Lambin X, Roholl P. Mycobacterium microti infection (vole tuberculosis) in wild rodent populations. Journal of Clinical Microbiology. 2002 Sep;40(9):3281-5.
  10. Sokhela MC. Development and validation of a molecular assay and evaluation of the GeneXpert® MTB/RIF assay for the rapid detection of genital tuberculosis (Doctoral dissertation, University of the Free State).
  11. Ryan GJ, Shapiro HM, Lenaerts AJ. Improving acid-fast fluorescent staining for the detection of mycobacteria using a new nucleic acid staining approach. Tuberculosis. 2014 Sep 1;94(5):511-8.
  12. Bunsow E, Ruiz-Serrano MJ, Roa PL, Kestler M, Viedma DG, Bouza E. Evaluation of GeneXpert MTB/RIF for the detection of Mycobacterium tuberculosis and resistance to rifampin in clinical specimens. Journal of Infection. 2014 Apr 1;68(4):338-43.
  13. Wang S, Duan H, Zhang W, Li JW. Analysis of bacterial foodborne disease outbreaks in China between 1994 and 2005. FEMS Immunology & Medical Microbiology. 2007 Oct 1;51(1):8-13.
  14. Kim YW, Seong MW, Kim TS, Yoo CG, Han SK, Yim JJ. Evaluation of Xpert® MTB/RIF assay: diagnosis and treatment outcomes in rifampicin-resistant tuberculosis. The International Journal of Tuberculosis and Lung Disease. 2015 Oct 1;19(10):1216-21.
  15. Neelakantan S, Nair PP, Emmanuel RV, Agrawal K. Diversities in presentations of extrapulmonary tuberculosis. Case Reports. 2013 Feb 28;2013: bcr2013008597.
  16. Maynard-Smith L, Larke N, Peters JA, Lawn SD. Diagnostic accuracy of the Xpert MTB/RIF assay for extrapulmonary and pulmonary tuberculosis when testing non-respiratory samples: a systematic review. BMC infectious diseases. 2014 Dec;14(1):1-5.
  17. Boyles TH, Hughes J, Cox V, Burton R, Meintjes G, Mendelson M. False-positive Xpert® MTB/RIF assays in previously treated patients: need for caution in interpreting results. The International journal of tuberculosis and lung disease. 2014 Jul 1;18(7):876-8.
  18. Alexander KA, Laver PN, Michel AL, Williams M, van Helden PD, Warren RM, van Pittius NC. Novel Mycobacterium tuberculosis complex pathogen, M. mungi. Emerging infectious diseases. 2010 Aug;16(8):1296. 
  19. al-Kassimi FA, al-Hajjaj MS, al-Orainey IO, Bamgboye EA. Does the protective effect of neonatal BCG correlate with vaccine-induced tuberculin reaction? American journal of respiratory and critical care medicine. 1995 Nov;152(5):1575-8.
  20. Alonso-Rodríguez N, Martínez-Lirola M, Herránz M, Sanchez-Benitez M, Barroso P, Bouza E, de Viedma DG. Evaluation of the new advanced 15-loci MIRU-VNTR genotyping tool in Mycobacterium tuberculosis molecular epidemiology studies. BMC microbiology. 2008 Dec;8(1):1-9.

 

 

 

 

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