Document Type : Original Article
Ministry of Science and Technology, Environment and water Directorate, Baghdad, Iraq
Department of Biomedical technologies, College of Biotechnology, Al-Nahrain University, Baghdad, Iraq.
Department of Biotechnology, Baghdad University, College of Medicine, Baghdad, Iraq
Background: Tuberculosis (TB) is considered to be one of the deadliest bacterial infections in the world and it can have a large impact on global health, drawing international attention with increasing number of cases worldwide in developed and developing countries. Mycobacterium tuberculosis (MTB) is the main causative agent of TB which is an aerobic pathogenic bacillus that establishes infection in the lungs. It is believed that two billion people are carrying non-eradicated intra-granulomatous Tuberculosis bacilli as LTBI (short for latent tuberculosis infection) and around 10% of those people will be infected with active tuberculosis during their lifetime. Therefore, this study aimed to study Mycobacterium tuberculosis in Iraq.
Methods: All bacteriological tests were done in biological safety cabinet class II. Gloves and masks (N 95) were used during the laboratory work. Phenol 5% and ethanol alcohol 70% were used to sterilize and disinfect the benches and hood (IUATLD, 1996). Autoclave was used at 121 °C at 1.5 bar for 20-30 minutes for sterilizing all culture media. The study was conducted at National Reference Lab (NRL) of tuberculosis/Baghdad from July 2016 to December 2016. A total of 188 sputum samples were collected from suspected patient. In addition, 80 samples were collected from healthy subjects Specimens were amassed within the outdoor and as away as feasible from different human beings. Each subject was advised to inhale deeply 2-3 instances, cough out deep from the chest and spit the sputum into sterile prevalent container. We needed to ensure that the specimen was of enough extent (3 to 5 mL) and that it incorporated strong or purulent material. Two samples were gathered from the subject. The first was taken when they reached the institute and the second was taken early morning, earlier than breakfast. The early morning collection represents the pulmonary secretions accrued in a single day, and consequently it typically has a better positivity. Gathered specimens were stored at –20 °C until use.
Results: Totally, 118 Tuberculosis suspected samples were used throughout this study; 103 (54.78%) cases were positive using direct assay (AFB smear microscopy) and 85 (45.21%) were negative cases. From the 103 TB patients, 88 (85.43%) represented as new TB cases; the others 15 (14.57%) patients appeared to follow up assessment of the directly observed treatment (DOT) program. The percentage of infected patients was 73 (70.87%) for males to 30 (29.12 %) for females, with over all male to female ratio of 2.43 (73/30) with a highly significant difference (P≤0.01). The age of the study patients ranged from 14 years to 75 years. The mean age was 36 and the median was 30 with highly significant difference between age group. As in many developing countries, direct sputum microscopy is the widely used method for the diagnosis of pulmonary TB in Iraq. Only 103 from 188 patients (suspected) were positive by using ZN smears (Ziehl-Neelsen stain technique). This study for those 188 suspected showed that 119 (63.29%) cases were positive by culture (Lowenstein–Jensen medium) and 69 (36.7%) cases were negative by L.J Culture. This study showed that the percentage of sensitivity, specificity, positive predictive value, and negative predictive value was 74.78%, 79.7%, 86.4% and 64.7%, respectively, using AFB method. The results revealed out of the total 188 specimens, 49 (26.06%) of the specimens were positive by DiaSpot (Tuberculosis Rapid Test on serum) and 139 (73.93%) specimens were negative. This study reported the sensitivity of 33.6% and the specificity of 86.95 % PPV 81.6 and NPV 43.16 for DiaSpot TB Rapid test.
Conclusion: Males aged 15 to 45 years are at high risk for TB infection. M. tuberculosis plays a major role for causing tuberculosis in human in Baghdad.
- Organization W H. (2014). Global tuberculosis report 2014: World Health Organization. 171 Pages,
- Rocchetti T T, Silbert S, Gostnell A, Kubasek C, Widen R. (2016). Validation of a multiplex real-time PCR assay for detection of Mycobacterium spp., Mycobacterium tuberculosis complex, and Mycobacterium avium complex directly from clinical samples by use of the BD Max open system. Journal of Clinical Microbiology, 54(6): 1644-1647. https://doi.org/10.1128/JCM.00241-16
- Moure R, Muñoz L, Torres M, Santin M, Martín R, Alcaide F. (2011). Rapid detection of Mycobacterium tuberculosis complex and rifampin resistance in smear-negative clinical samples by use of an integrated real-time PCR method. Journal of Clinical Microbiology, 49(3): 1137-1139. https://doi.org/10.1128/JCM.01831-10
- Zakham F, Akrim M, Benjouad A, Ennaji M. (2012). Rapid screening and diagnosis of tuberculosis: a real challenge for the mycobacteriologist. Cellular and Molecular Biology, 58(2): 1632-1640.
- Yusuf E. (2015). A Book Review on: Sherris Medical Microbiology–International Edition. Frontiers in cellular and infection microbiology, 5: 34. https://doi.org/10.3389/fcimb.2015.00034
- LoBue P, Enarson D, Thoen C. (2010). Tuberculosis in humans and animals: an overview [Serialised article. Tuberculosis: a re-emerging disease in animals and humans. Number 1 in the series]. The International Journal of Tuberculosis and Lung Disease, 14(9): 1075-1078.
- Chakaya J, Khan M, Ntoumi F, Aklillu E, Fatima R, Mwaba P, Kapata N, Mfinanga S, Hasnain S E, Katoto P D. (2021). Global Tuberculosis Report 2020–Reflections on the Global TB burden, treatment and prevention efforts. International Journal of Infectious Diseases, 113: S7-S12. https://doi.org/10.1016/j.ijid.2021.02.107
- Corbett E L, Watt C J, Walker N, Maher D, Williams B G, Raviglione M C, Dye C. (2003). The growing burden of tuberculosis: global trends and interactions with the HIV epidemic. Archives of Internal Medicine, 163(9): 1009-1021. https://doi.org/10.1001/archinte.163.9.1009
- Hopewell P C, Fair E L, Uplekar M. (2014). Updating the International Standards for Tuberculosis Care. Entering the era of molecular diagnostics. Annals of the American Thoracic Society, 11(3): 277-285. https://doi.org/10.1513/AnnalsATS.201401-004AR
- Karabulut F, Parray J A, Mir M Y. (2021). Emerging trends for Harnessing plant metabolome and microbiome for sustainable food Production. Micro Environer, 1(1): 33-53 https://doi.org/10.54458/mev.v1i01.6672
- Abbas-Al-Khafaji Z-K, Aubais-aljelehawy Q h. (2021). Evaluation of antibiotic resistance and prevalence of multi-antibiotic resistant genes among Acinetobacter baumannii strains isolated from patients admitted to al-yarmouk hospital. Cellular, Molecular and Biomedical Reports, 1(2): 60-68.
- Hutchins R J, Phan K L, Saboor A, Miller J D, Muehlenbachs A. (2019). Practical guidance to implementing quality management systems in public health laboratories performing next-generation sequencing: personnel, equipment, and process management (phase 1). Journal of Clinical Microbiology, 57(8): e00261-00219. https://doi.org/10.1128/JCM.00261-19
- Robinson S. (2021). Priorities for Health Promotion and Public Health: Londen: Routledge. 111 Pages,
- Subgroup S T P C T. (2006). Guidance for National Tuberculosis Programmes on the management of tuberculosis in children. Chapter 1: introduction and diagnosis of tuberculosis in children. The international journal of tuberculosis and lung disease: the official journal of the International Union against Tuberculosis and Lung Disease, 10(10): 1091-1097. PMID: 17044200
- Mankhi A. (2009). Supervised by Hashem, ZS Guidelines of Laboratory manual work of tuberculosis in Iraq. Iraq. Baghdad: Prepared by Ministry of Health. 289 Pages,
- Mostafa H K, Risan M H, Al-Faham M, Tbena M. (2018). Detection of Mycobacterium tuberculosis from Clinical Samples using Different Real-Time PCR System. Int. J. Curr. Microbiol. App. Sci, 7(12): 2357-2368. https://doi.org/10.20546/ijcmas.2018.712.267
- Al-Saadi M, Naher H, Shalan A. (2014). Evaluation of Commercial Real Time PCR and Immunochromatography Techniques in Laboratory Diagnosis of Tuberculosis. Medical Journal of Babylon, 11(1): 58-69.
- Rosser E, John Holley M, Choi H, Harvey S A. (2009). Willingness to use and pay for a new diagnostic test for active tuberculosis in HIV-positive and HIV-negative individuals: Results from Benin, Peru, and Tanzania. Bethesda: Center for Human Services. 71 Pages,
- Parray J A, Ali U, Mir M Y, Shameem N. (2021). A high throughputs and consistent method for the sampling and isolation of Endophytic bacteria allied to high altitude the medicinal plant Arnebia benthamii (Wall ex. G. Don). Micro Environer, 1(1): 1-6. https://doi.org/10.54458/mev.v1i01.6668
- Duyan V, Kurt B, Aktas Z, Duyan G, Kulkul D. (2005). Relationship between quality of life and characteristics of patients hospitalised with tuberculosis. The International Journal of Tuberculosis and Lung Disease, 9(12): 1361-1366.
- Djibuti M, Mirvelashvili E, Makharashvili N, Magee M J. (2014). Household income and poor treatment outcome among patients with tuberculosis in Georgia: a cohort study. BMC Public Health, 14(1): 1-7. https://doi.org/10.1186/1471-2458-14-88
- Mohammed S H, Ahmed M M, Ahmed A R H. (2013). First experience with using simple polymerase chain reaction-based methods as an alternative to phenotypic drug susceptibility testing for Myobacterium tuberculosis in Iraq. International Journal of Applied and Basic Medical Research, 3(2): 98. https://dx.doi.org/10.4103%2F2229-516X.117069
- Shaker H, Saleh D. (2013). A study on diagnosis and multidrug resistance of Mycobacterium tuberculosis using different methods. Unpublished thesis for the Degree of Master of Science in Microbiology. College of Science/University of Baghdad.
- Boum Y, Atwine D, Orikiriza P, Assimwe J, Page A-L, Mwanga-Amumpaire J, Bonnet M. (2014). Male Gender is independently associated with pulmonary tuberculosis among sputum and non-sputum producers people with presumptive tuberculosis in Southwestern Uganda. BMC infectious diseases, 14(1): 1-8. https://doi.org/10.1186/s12879-014-0638-5
- Babamahmoodi F, Alikhani A, Yazdani Charati J, Ghovvati A, Ahangarkani F, Delavarian L, Babamahmoodi A. (2015). Clinical epidemiology and paraclinical findings in tuberculosis patients in north of Iran. BioMed research international, 2015(Article ID 381572): 1-5. https://doi.org/10.1155/2015/381572
- Roy A, Eisenhut M, Harris R, Rodrigues L, Sridhar S, Habermann S, Snell L, Mangtani P, Adetifa I, Lalvani A. (2014). Effect of BCG vaccination against Mycobacterium tuberculosis infection in children: systematic review and meta-analysis. Bmj, 349: g4643 https://doi.org/10.1136/bmj.g4643
- Singh P N, Yel D, Hurd G, Job J S. (2013). Cigarette smoking and tuberculosis in Cambodia: findings from a national sample. Tobacco Induced Diseases, 11(1): 1-9. https://doi.org/10.1186/1617-9625-11-8
- Al-Jebouri M M, Wahid N M. (2014). The interactions between polymerase chain reaction (PCR) and other diagnostic tests among tuberculosis patients of various blood groups. World J. pharm. Pharmaceut. sci, 3(7): 217-230.
- Yarahmadi A, Zahmatkesh M M, Ghaffari M, Mohammadi S, Labbafinejad Y, Seyedmehdi S M, Nojomi M, Attarchi M. (2013). Correlation between silica exposure and risk of tuberculosis in Lorestan Province of Iran. Tanaffos, 12(2): 34. PMCID: PMC4153244; PMID: 25191460
- Rahmo A, Hamze M. (2010). Characterization of Mycobacterium tuberculosis in Syrian patients by double-repetitive-element polymerase chain reaction. EMHJ-Eastern Mediterranean Health Journal, 16 (8), 820-830, 2010, 16(8): 820-830.
- Khosravi A D, Dezfulian A, Alavi S M. (2006). Detection of Isoniazid and Rifampin resistant Mycobacterium tuberculosis isolated from tuberculosis patients using conventional method and PCR. PAKISTAN JOURNAL OF MEDICAL SCIENCES, 22(1): 47-52.
- Sharma S K, Kaushik G, Jha B, George N, Arora S, Gupta D, Singh U, Hanif M, Vashisht R. (2011). Prevalence of multidrug-resistant tuberculosis among newly diagnosed cases of sputum-positive pulmonary tuberculosis. The Indian Journal of Medical Research, 133(3): 308. PMCID: PMC3103156; PMID: 21441685
- Schaaf H S, Collins A, Bekker A, Davies P D. (2010). Tuberculosis at extremes of age. Respirology, 15(5): 747-763. https://doi.org/10.1111/j.1440-1843.2010.01784.x
- Datta S, Sherman J M, Bravard M A, Valencia T, Gilman R H, Evans C A. (2015). Clinical evaluation of tuberculosis viability microscopy for assessing treatment response. Clinical infectious diseases, 60(8): 1186-1195. https://doi.org/10.1093/cid/ciu1153
- Chan C E, Zhao B Z, Cazenave-Gassiot A, Pang S-W, Bendt A K, Wenk M R, MacAry P A, Hanson B J. (2013). Novel phage display-derived mycolic acid-specific antibodies with potential for tuberculosis diagnosis [S]. Journal of lipid research, 54(10): 2924-2932. https://doi.org/10.1194/jlr.D036137
- Udwadia Z F. (2003). Controlling tuberculosis in India. The New England journal of medicine, 348(8): 758-759; author reply 758. https://doi.org/10.1056/nejm200302203480817
- Organization W H. (2006). Diagnostic and treatment delay in tuberculosis. https://apps.who.int/iris/handle/10665/116501.
- Kassa-Kelembho E, Kassa E, Zandanga G, Service Y-B, Ignaleamoko A, Talarmin A. (2006). Poor performance of a novel serological test for diagnosis of pulmonary tuberculosis in Bangui, Central African Republic. Clinical and vaccine immunology, 13(6): 702-703. https://doi.org/10.1128/CVI.00194-05
- Steingart K R, Henry M, Laal S, Hopewell P C, Ramsay A, Menzies D, Cunningham J, Weldingh K, Pai M. (2007). Commercial serological antibody detection tests for the diagnosis of pulmonary tuberculosis: a systematic review. PLoS medicine, 4(6): e202. https://doi.org/10.1371/journal.pmed.0040202
- Sada E, Aguilar D, Torres M, Herrera T. (1992). Detection of lipoarabinomannan as a diagnostic test for tuberculosis. Journal of Clinical Microbiology, 30(9): 2415-2418. https://doi.org/10.1128/jcm.30.9.2415-2418.1992
- Davidow A, Kanaujia G V, Shi L, Kaviar J, Guo X, Sung N, Kaplan G, Menzies D, Gennaro M L. (2005). Antibody profiles characteristic of Mycobacterium tuberculosis infection state. Infection and immunity, 73(10): 6846-6851. https://doi.org/10.1128/IAI.73.10.6846-6851.2005