Time to detection of Mycobacterium tuberculosis using culture filtrate H37rv supplementation on MGIT 960 System

Resti Enggar Paweninggalih; Ni Made Mertaniasih; Eko Budi  Koendhori; Soedarsono Soedarsono

doi:10.15562/bmj.v12i1.3833

Time to detection of Mycobacterium tuberculosis using culture filtrate H37rv supplementation on MGIT 960 System

Resti Enggar Paweninggalih ,

Ni Made Mertaniasih ,

Eko Budi Koendhori ,

Soedarsono Soedarsono ,

pdf |
DOI: https://doi.org/10.15562/bmj.v12i1.3833 |
Published: 2023-01-07

Abstract

Background: The fundamental priority program for controlling Tuberculosis (TB) is accurate and rapid diagnosis. Variable Mycobacterium tuberculosis (MTB) phenotypes were discovered in sputum samples from pulmonary TB (PTB) patients. Active replicating bacteria and differentially culturable tubercle bacteria (DCTB) are present in the sputum. Differentially culturable tubercle bacilli did not grow on the standard medium. Culture filtrate (CF) H37Rv supplementation containing resuscitation-promoting factors (Rpfs) has the potential to increase sensitivity and reduce time to detection (TTD) on culture-based diagnostics.

Methods: This study was a true experimental laboratory. A clinical sample was taken from the sputum of 15 PTB patients, which was "MTB detected" by Xpert MTB/RIF assay. Sputum samples were divided into two groups and inoculated on MGIT 960 in one group with CF-H37Rv supplementation and without in the other as standard MGIT medium.

Results: On both standard MGIT medium and MGIT+CF H37Rv, the recovery rate was 100%. Seven samples (47%) show a growth promotion effect with CF H37Rv supplementation (ΔTTD > 10 hours), while eight samples (53%) show no change in TTD (ΔTTD < 10 hours). There is no evidence that CF H37Rv supplementation inhibits growth. Six of the seven samples with ΔTTD > 10 jam (85,7%) were dominated by samples with acid-fast bacilli (AFB) grading 1+, which could be an assumption related to re-treatment PTB cases, chronic cases, treatment stages, TB-HIV, or follow up of treatments.

Conclusion: There was no difference in recovery rate or TTD between standard MGIT medium and MGIT+CF H37Rv. The supplementation of culture filtrate H37Rv has a greater advantage on samples with low bacterial load (AFB 1+).

References

World Health Organization (WHO). Global Tuberculosis Report 2021. Geneva, Switzerland. 2021. [Cited 26th September 2022] [Available from: https://www.who.int/publications/i/item/9789240037021]
World Health Organization (WHO). Global Tuberculosis Report 2020. Geneva: WHO. 2020. [Cited 26th September 2022] [Available from: https://www.who.int/publications/i/item/9789240013131]
Pai M, Nicol MP, Boehme CC. Tuberculosis Diagnostics: State of the Art and Future Directions. Microbiol Spectr. 2016;4(5):1-15.
World Health Organization (WHO). Regional strategic plan towards ending TB in WHO Southeast-Asia region:2021-2025. India: WHO press. 2021. [Cited 26th September 2022] [Available from: https://www.who.int/publications/i/item/9789290228974]
Dharmadhikari AS, Mphahlele M, Venter K, Stoltz A, Mathebula R, Masotla T, et al. Rapid impact of effective treatment on transmission of multidrug-resistant tuberculosis. Int J Tuberc Lung Dis. 2014;18(9):1019–1025.
Parrish NM, Carroll KC. Role of the clinical mycobacteriology laboratory in diagnosis and management of tuberculosis in low-prevalence settings. J Clin Microbiol. 2011;49(3):772–776.
Viveiros M, Machado D, Couto I, Amaral L. Improving on the LJ slope—Automated liquid culture. Diagnosis and Treatment, Advances in Molecular and Cellular Microbiology Series. McHugh. 2013:34–45.
Chengalroyen MD, Beukes GM, Gordhan BG, Streicher EM, Churchyard G, Hafner R, et al. Detection and Quantification of Differentially Culturable Tubercle Bacteria in Sputum from Patients with Tuberculosis. Am J Respir Crit Care Med. 2016;194(12):1532–1540.
Mukamolova GV, Turapov O, Malkin J, Woltmann G, Barer MR. Resuscitation-promoting factors reveal an occult population of tubercle Bacilli in Sputum. Am J Respir Crit Care Med. 2010;181(2):174–180.
Beltran CGG, Heunis T, Gallant J, Venter R, du Plessis N, Loxton AG, et al. Investigating Non-sterilizing Cure in TB Patients at the End of Successful Anti-TB Therapy. Front Cell Infect Microbiol. 2020;10:443.
Rosser A, Stover C, Pareek M, Mukamolova G V. Resuscitation-promoting factors are important determinants of the pathophysiology in Mycobacterium tuberculosis infection. Crit Rev Microbiol. 2017;43(5):621–630.
Peddireddy V, Doddam SN, Ahmed N. Mycobacterial Dormancy Systems and Host Responses in Tuberculosis. Front Immunol. 2017;8:1–19.
Dusthackeer A, Balasubramanian M, Shanmugam G, Priya S, Nirmal CR, Sam Ebenezer R, et al. Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy. Front Microbiol. 2019;10:2381.
Kana BD, Gordhan BG, Downing KJ, Sung N, Vostroktunova G, Machowski EE, et al. The resuscitation-promoting factors of Mycobacterium tuberculosis are required for virulence and resuscitation from dormancy but are collectively dispensable for growth in vitro. Mol Microbiol. 2008;67(3):672–684.
Kana BD, Mizrahi V. Resuscitation-promoting factors as lytic enzymes for bacterial growth and signaling. FEMS Immunol Med Microbiol. 2010;58(1):39–50.
Dworkin J, Shah IM. Exit from dormancy in microbial organisms. Nat Rev Microbiol. 2010;8(12):890–896.
Kolwijck E, Friedrich SO, Karinja MN, van Ingen J, Warren RM, Diacon AH. Early stationary phase culture supernatant accelerates the growth of sputum cultures collected after initiation of anti-tuberculosis treatment. Clin Microbiol Infect. 2014;20(7):O418-O420.
Maitra A, Munshi T, Healy J, Martin LT, Vollmer W, Keep NH, et al. Cell wall peptidoglycan in Mycobacterium tuberculosis: An Achilles’ heel for the TB-causing pathogen. FEMS Microbiol Rev. 2019;43(5):548–575.
Mukamolova GV, Turapov OA, Young DI, Kaprelyants AS, Kell DB, Young M. A family of autocrine growth factors in Mycobacterium tuberculosis. Mol Microbiol. 2002;46(3):623–635.
Gupta RK, Srivastava R. Resuscitation Promoting Factors: a Family of Microbial Proteins in Survival and Resuscitation of Dormant Mycobacteria. Indian J Microbiol. 2012;52(2):114–121.
Shleeva MO, Bagramyan K, Telkov MV, Mukamolova GV, Young M, Kell DB, et al. Formation and resuscitation of “non-culturable” cells of Rhodococcus rhodochrous and Mycobacterium tuberculosis in prolonged stationary phase. Microbiology. 2002;148(5):1581–1591.
Stinson K, Eisenach K, Kayes S, Matsumoto M, Siddiqi S, Nakashima S, et al. Global Laboratory Initiative. 2014. [Cited 20th October 2022] [Available from: https://stoptb.org/wg/gli/assets/documents/gli_mycobacteriology_lab_manual_web.pdf].
Diriba G, Kebede A, Yaregal Z, Getahun M, Tadesse M, Meaza A, et al. Performance of Mycobacterium Growth Indicator Tube BACTEC 960 with Lowenstein-Jensen method for diagnosis of Mycobacterium tuberculosis at Ethiopian National Tuberculosis Reference Laboratory, Addis Ababa, Ethiopia. BMC Res Notes. 2017;10(1):181.
Gordhan BG, Peters JS, McIvor A, Machowski EE, Ealand C, Waja Z, et al. Detection of differentially culturable tubercle bacteria in sputum using mycobacterial culture filtrates. Sci Rep. 2021;11(1):6493.
McIvor A, Gordhan BG, Waja Z, Otwombe K, Martinson NA, Kana BD. Supplementation of sputum cultures with culture filtrate to detect tuberculosis in a cross-sectional study of HIV-infected individuals. Tuberculosis (Edinb). 2021;129:102103.
Peñuelas-Urquides K, Villarreal-Treviño L, Silva-Ramírez B, Rivadeneyra-Espinoza L, Said-Fernández S, León MBD. Measuring of Mycobacterium tuberculosis growth: a correlation of the optical measurements with colony forming units. Brazilian J Microbiol. 2013;44(1):287–290.
Siddiqi SH, Rusch-Gerdes S. BACTEC MGIT 960 System for mycobacteria testing. BD diagnostic system. Beckton Dickinson. 2006. [Cited 13th October 2022] [Available from: https://www.finddx.org/wp-content/uploads/2016/02/mgit_manual_nov2006.pdf]
Kementerian Kesehatan Republik Indonesia. National Guidelines for Tuberculosis Management Medical Services. Jakarta: Health Ministry of Indonesia. 2020. [Cited 20th October 2022] [Available from: https://tbindonesia.or.id/pustaka/pedoman/umum/pedoman-nasional-pelayanan-kedokteran-tata-laksana-tuberkulosis/]
Bhat J, Rao V, Muniyandi M, Yadav R, Karforma C, Luke C. Impact of sputum quality and quantity on smear and culture positivity: findings from a tuberculosis prevalence study in central India. Trans R Soc Trop Med Hyg. 2014;108(1):55–56.
Huang W, Qi Y, Diao Y, Yang F, Zha X, Ren C, et al. use of resuscitation-promoting factor proteins improves the sensitivity of culture-based tuberculosis testing in special samples. Am J Respir Crit Care Med 2014;189(5):612–614.
Yoon SH, Lee NK, Yim JJ. Impact of sputum gross appearance and volume on smear positivity of pulmonary tuberculosis: a prospective cohort study. BMC Infect Dis. 2012;12(1):172.
McAulay K, Saito K, Warrier T, Walsh KF, Mathurin LD, Royal-Mardi G, et al. Differentially Detectable Mycobacterium tuberculosis Cells in Sputum from Treatment-Naive Subjects in Haiti and Their Proportionate Increase after Initiation of Treatment. MBio. 2018;9(6):1–12.
Sarathy JP, Via LE, Weiner D, Blanc L, Boshoff H, Eugenin EA, et al. Extreme Drug Tolerance of Mycobacterium tuberculosis in Caseum. Antimicrob Agents Chemother. 2018;62(2):1–11.
Turapov O, Waddell SJ, Burke B, Glenn S, Sarybaeva AA, Tudo G, et al. Antimicrobial treatment improves mycobacterial survival in nonpermissive growth conditions. Antimicrob Agents Chemother. 2014;58(5):2798–2806.
Almeida Júnior PS, Schmidt Castellani LG, Peres RL, Combadao J, Tristão TC, Dietze R, et al. Differentially culturable tubercle bacteria dynamics during standard anti-tuberculosis treatment: A prospective cohort study. Tuberculosis (Edinb). 2020;124:101945.
Gordhan BG, Sewcharran A, Letsoalo M, Chinappa T, Yende-Zuma N, Padayatchi N, et al. Detection of differentially culturable tubercle bacteria in sputum from drug-resistant tuberculosis patients. Front Cell Infect Microbiol. 2022;12:949370.
Dewi DNSS, Soedarsono S, Kurniati A, Mertaniasih NM. The specific DNA region of esxA gene for the target of PCR to determine Mycobacterium tuberculosis accurately. Bali Medical Journal. 2017;6(1):150-155.
Herawati S, Kandarini Y, Mulyantari NK, Prabawa PY. Correlation of Neutrophil to Lymphocyte Ratio with Interleukin-10 in Diagnosis and Monitoring of Coronavirus Disease-19 Patients. Open Access Maced J Med Sci. 2022;10(B):63-66.

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[2] World Health Organization (WHO). Global Tuberculosis Report 2020. Geneva: WHO. 2020. [Cited 26th September 2022] [Available from: https://www.who.int/publications/i/item/9789240013131]

[3] Pai M, Nicol MP, Boehme CC. Tuberculosis Diagnostics: State of the Art and Future Directions. Microbiol Spectr. 2016;4(5):1-15.

[4] World Health Organization (WHO). Regional strategic plan towards ending TB in WHO Southeast-Asia region:2021-2025. India: WHO press. 2021. [Cited 26th September 2022] [Available from: https://www.who.int/publications/i/item/9789290228974]

[5] Dharmadhikari AS, Mphahlele M, Venter K, Stoltz A, Mathebula R, Masotla T, et al. Rapid impact of effective treatment on transmission of multidrug-resistant tuberculosis. Int J Tuberc Lung Dis. 2014;18(9):1019–1025.

[6] Parrish NM, Carroll KC. Role of the clinical mycobacteriology laboratory in diagnosis and management of tuberculosis in low-prevalence settings. J Clin Microbiol. 2011;49(3):772–776.

[7] Viveiros M, Machado D, Couto I, Amaral L. Improving on the LJ slope—Automated liquid culture. Diagnosis and Treatment, Advances in Molecular and Cellular Microbiology Series. McHugh. 2013:34–45.

[8] Chengalroyen MD, Beukes GM, Gordhan BG, Streicher EM, Churchyard G, Hafner R, et al. Detection and Quantification of Differentially Culturable Tubercle Bacteria in Sputum from Patients with Tuberculosis. Am J Respir Crit Care Med. 2016;194(12):1532–1540.

[9] Mukamolova GV, Turapov O, Malkin J, Woltmann G, Barer MR. Resuscitation-promoting factors reveal an occult population of tubercle Bacilli in Sputum. Am J Respir Crit Care Med. 2010;181(2):174–180.

[10] Beltran CGG, Heunis T, Gallant J, Venter R, du Plessis N, Loxton AG, et al. Investigating Non-sterilizing Cure in TB Patients at the End of Successful Anti-TB Therapy. Front Cell Infect Microbiol. 2020;10:443.

[11] Rosser A, Stover C, Pareek M, Mukamolova G V. Resuscitation-promoting factors are important determinants of the pathophysiology in Mycobacterium tuberculosis infection. Crit Rev Microbiol. 2017;43(5):621–630.

[12] Peddireddy V, Doddam SN, Ahmed N. Mycobacterial Dormancy Systems and Host Responses in Tuberculosis. Front Immunol. 2017;8:1–19.

[13] Dusthackeer A, Balasubramanian M, Shanmugam G, Priya S, Nirmal CR, Sam Ebenezer R, et al. Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy. Front Microbiol. 2019;10:2381.

[14] Kana BD, Gordhan BG, Downing KJ, Sung N, Vostroktunova G, Machowski EE, et al. The resuscitation-promoting factors of Mycobacterium tuberculosis are required for virulence and resuscitation from dormancy but are collectively dispensable for growth in vitro. Mol Microbiol. 2008;67(3):672–684.

[15] Kana BD, Mizrahi V. Resuscitation-promoting factors as lytic enzymes for bacterial growth and signaling. FEMS Immunol Med Microbiol. 2010;58(1):39–50.

[16] Dworkin J, Shah IM. Exit from dormancy in microbial organisms. Nat Rev Microbiol. 2010;8(12):890–896.

[17] Kolwijck E, Friedrich SO, Karinja MN, van Ingen J, Warren RM, Diacon AH. Early stationary phase culture supernatant accelerates the growth of sputum cultures collected after initiation of anti-tuberculosis treatment. Clin Microbiol Infect. 2014;20(7):O418-O420.

[18] Maitra A, Munshi T, Healy J, Martin LT, Vollmer W, Keep NH, et al. Cell wall peptidoglycan in Mycobacterium tuberculosis: An Achilles’ heel for the TB-causing pathogen. FEMS Microbiol Rev. 2019;43(5):548–575.

[19] Mukamolova GV, Turapov OA, Young DI, Kaprelyants AS, Kell DB, Young M. A family of autocrine growth factors in Mycobacterium tuberculosis. Mol Microbiol. 2002;46(3):623–635.

[20] Gupta RK, Srivastava R. Resuscitation Promoting Factors: a Family of Microbial Proteins in Survival and Resuscitation of Dormant Mycobacteria. Indian J Microbiol. 2012;52(2):114–121.

[21] Shleeva MO, Bagramyan K, Telkov MV, Mukamolova GV, Young M, Kell DB, et al. Formation and resuscitation of “non-culturable” cells of Rhodococcus rhodochrous and Mycobacterium tuberculosis in prolonged stationary phase. Microbiology. 2002;148(5):1581–1591.

[22] Stinson K, Eisenach K, Kayes S, Matsumoto M, Siddiqi S, Nakashima S, et al. Global Laboratory Initiative. 2014. [Cited 20th October 2022] [Available from: https://stoptb.org/wg/gli/assets/documents/gli_mycobacteriology_lab_manual_web.pdf].

[23] Diriba G, Kebede A, Yaregal Z, Getahun M, Tadesse M, Meaza A, et al. Performance of Mycobacterium Growth Indicator Tube BACTEC 960 with Lowenstein-Jensen method for diagnosis of Mycobacterium tuberculosis at Ethiopian National Tuberculosis Reference Laboratory, Addis Ababa, Ethiopia. BMC Res Notes. 2017;10(1):181.

[24] Gordhan BG, Peters JS, McIvor A, Machowski EE, Ealand C, Waja Z, et al. Detection of differentially culturable tubercle bacteria in sputum using mycobacterial culture filtrates. Sci Rep. 2021;11(1):6493.

[25] McIvor A, Gordhan BG, Waja Z, Otwombe K, Martinson NA, Kana BD. Supplementation of sputum cultures with culture filtrate to detect tuberculosis in a cross-sectional study of HIV-infected individuals. Tuberculosis (Edinb). 2021;129:102103.

[26] Peñuelas-Urquides K, Villarreal-Treviño L, Silva-Ramírez B, Rivadeneyra-Espinoza L, Said-Fernández S, León MBD. Measuring of Mycobacterium tuberculosis growth: a correlation of the optical measurements with colony forming units. Brazilian J Microbiol. 2013;44(1):287–290.

[27] Siddiqi SH, Rusch-Gerdes S. BACTEC MGIT 960 System for mycobacteria testing. BD diagnostic system. Beckton Dickinson. 2006. [Cited 13th October 2022] [Available from: https://www.finddx.org/wp-content/uploads/2016/02/mgit_manual_nov2006.pdf]

[28] Kementerian Kesehatan Republik Indonesia. National Guidelines for Tuberculosis Management Medical Services. Jakarta: Health Ministry of Indonesia. 2020. [Cited 20th October 2022] [Available from: https://tbindonesia.or.id/pustaka/pedoman/umum/pedoman-nasional-pelayanan-kedokteran-tata-laksana-tuberkulosis/]

[29] Bhat J, Rao V, Muniyandi M, Yadav R, Karforma C, Luke C. Impact of sputum quality and quantity on smear and culture positivity: findings from a tuberculosis prevalence study in central India. Trans R Soc Trop Med Hyg. 2014;108(1):55–56.

[30] Huang W, Qi Y, Diao Y, Yang F, Zha X, Ren C, et al. use of resuscitation-promoting factor proteins improves the sensitivity of culture-based tuberculosis testing in special samples. Am J Respir Crit Care Med 2014;189(5):612–614.

[31] Yoon SH, Lee NK, Yim JJ. Impact of sputum gross appearance and volume on smear positivity of pulmonary tuberculosis: a prospective cohort study. BMC Infect Dis. 2012;12(1):172.

[32] McAulay K, Saito K, Warrier T, Walsh KF, Mathurin LD, Royal-Mardi G, et al. Differentially Detectable Mycobacterium tuberculosis Cells in Sputum from Treatment-Naive Subjects in Haiti and Their Proportionate Increase after Initiation of Treatment. MBio. 2018;9(6):1–12.

[33] Sarathy JP, Via LE, Weiner D, Blanc L, Boshoff H, Eugenin EA, et al. Extreme Drug Tolerance of Mycobacterium tuberculosis in Caseum. Antimicrob Agents Chemother. 2018;62(2):1–11.

[34] Turapov O, Waddell SJ, Burke B, Glenn S, Sarybaeva AA, Tudo G, et al. Antimicrobial treatment improves mycobacterial survival in nonpermissive growth conditions. Antimicrob Agents Chemother. 2014;58(5):2798–2806.

[35] Almeida Júnior PS, Schmidt Castellani LG, Peres RL, Combadao J, Tristão TC, Dietze R, et al. Differentially culturable tubercle bacteria dynamics during standard anti-tuberculosis treatment: A prospective cohort study. Tuberculosis (Edinb). 2020;124:101945.

[36] Gordhan BG, Sewcharran A, Letsoalo M, Chinappa T, Yende-Zuma N, Padayatchi N, et al. Detection of differentially culturable tubercle bacteria in sputum from drug-resistant tuberculosis patients. Front Cell Infect Microbiol. 2022;12:949370.

[37] Dewi DNSS, Soedarsono S, Kurniati A, Mertaniasih NM. The specific DNA region of esxA gene for the target of PCR to determine Mycobacterium tuberculosis accurately. Bali Medical Journal. 2017;6(1):150-155.

[38] Herawati S, Kandarini Y, Mulyantari NK, Prabawa PY. Correlation of Neutrophil to Lymphocyte Ratio with Interleukin-10 in Diagnosis and Monitoring of Coronavirus Disease-19 Patients. Open Access Maced J Med Sci. 2022;10(B):63-66.

Time to detection of Mycobacterium tuberculosis using culture filtrate H37rv supplementation on MGIT 960 System

Abstract

References

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