Journal of Microbiology and Biotechnology

  • 제24권8호
  • /
  • Pages.1044-1050
  • /
  • 2014
  • /
  • 1017-7825(pISSN)
  • /
  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지
DOI QR코드

DOI QR Code

Evaluation of Galactomannan Enzyme Immunoassay and Quantitative Real-Time PCR for the Diagnosis of Invasive Pulmonary Aspergillosis in a Rat Model

  • Lin, Jian-Cong (Department of Internal Medicine, Huang Pu Hospital of the First Affiliated Hospital, Sun Yat-sen University) ;
  • Xing, Yan-Li (Department of Internal Medicine, Huang Pu Hospital of the First Affiliated Hospital, Sun Yat-sen University) ;
  • Xu, Wen-Ming (Department of Internal Medicine, Huang Pu Hospital of the First Affiliated Hospital, Sun Yat-sen University) ;
  • Li, Ming (Department of Internal Medicine, Huang Pu Hospital of the First Affiliated Hospital, Sun Yat-sen University) ;
  • Bo, Pang (Department of Internal Medicine, Huang Pu Hospital of the First Affiliated Hospital, Sun Yat-sen University) ;
  • Niu, Yuan-Yuan (Department of Internal Medicine, Huang Pu Hospital of the First Affiliated Hospital, Sun Yat-sen University) ;
  • Zhang, Chang-Ran (Department of Internal Medicine, Huang Pu Hospital of the First Affiliated Hospital, Sun Yat-sen University)
  • 투고 : 2014.02.10
  • 심사 : 2014.04.17
  • 발행 : 2014.08.28
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Since there is no consensus about the most reliable assays to detect invasive aspergillosis from samples obtained by minimally invasive or noninvasive methods, we compared the efficacy of an enzyme-linked immunosorbent assay (ELISA) for galactomannan (GM) detection and quantitative real-time PCR assay (qRT-PCR) for the diagnosis of invasive pulmonary aspergillosis. Neutropenic, male Sprague-Dawley rats (specific pathogen free; 8 weeks old; weight, $200{\pm}20g$) were immunosuppressed with cyclophosphamide and infected with Aspergillus fumigatus intratracheally. Tissue and whole blood samples were harvested on days 1, 3, 5, and 7 post-infection and examined with GM ELISA and qRT-PCR. The A. fumigatus DNA detection sequence was detected in the following number of samples from 12 immunosuppressed, infected rats examined on the scheduled days: day 1 (0/12), day 3 (0/12), day 5 (6/12), and day 7 (8/12) post-infection. The sensitivity and specificity of the qRT-PCR assay was 29.2% and 100%, respectively. Receiver operating characteristic curve (ROC) analysis indicated a Ct (cycle threshold) cut-off value of 15.35, and the area under the curve (AUC) was 0.627. The GM assay detected antigen in sera obtained on day 1 (5/12), day 3 (9/12), day 5 (12/12), and day 7 (12/12) post-infection, and thus had a sensitivity of 79.2% and a specificity of 100%. The ROC of the GM assay indicated that the optimal Ct cut-off value was 1.40 (AUC, 0.919). The GM assay was more sensitive than the qRT-PCR assay in diagnosing invasive pulmonary aspergillosis in rats.

키워드

참고문헌

  1. Aquino VR, Nagel F, Andreolla HF, de-Paris F, Xavier MO, Goldani LZ, et al. 2012. The performance of real-time PCR, galactomannan, and fungal culture in the diagnosis of invasive aspergillosis in ventilated patients with chronic obstructive pulmonary disease (COPD). Mycopathologia 174: 163-169. https://doi.org/10.1007/s11046-012-9531-1
  2. Becker MJ, de Marie S, Willemse D, Verbrugh HA, Bakker- Woudenberg IA. 2000. Quantitative galactomannan detection is superior to PCR in diagnosing and monitoring invasive pulmonary aspergillosis in an experimental rat model. J. Clin. Microbiol. 38: 1434-1438.
  3. Chamilos G, Luna M, Lewis RE, Bodey GP, Chemaly R, Tarrand JJ, et al. 2006. Invasive fungal infections in patients with hematologic malignancies in a tertiary care cancer center: an autopsy study over a 15-year period (1989-2003). Haematologica 91: 986-989.
  4. Groll AH, McNeil Grist L. 2009. Current challenges in the diagnosis and management of invasive fungal infections: report from the 15th International Symposium on Infections in the Immunocompromised Host: Thessaloniki, Greece, 22- 25 June 2008. Int. J. Antimicrob. Agents 33: 101-104. https://doi.org/10.1016/j.ijantimicag.2008.08.014
  5. Hadrich I, Makni F, Cheikhrouhou F, Neji S, Amouri I, Sellami H, et al. 2012. Clinical utility and prognostic value of galactomannan in neutropenic patients with invasive aspergillosis. Pathol. Biol. (Paris) 60: 357-361. https://doi.org/10.1016/j.patbio.2011.10.011
  6. Hadrich I, Mary C, Makni F, Elloumi M, Dumon H, Ayadi A, Ranque S. 2011. Comparison of PCR-ELISA and real-time PCR for invasive aspergillosis diagnosis in patients with hematological malignancies. Med. Mycol. 49: 489-494.
  7. Hahn-Ast C, Glasmacher A, Muckter S, Schmitz A, Kraemer A, Marklein G, et al. 2010. Overall survival and fungal infection-related mortality in patients with invasive fungal infection and neutropenia after myelosuppressive chemotherapy in a tertiary care centre from 1995 to 2006. J. Antimicrob. Chemother. 65: 761-768. https://doi.org/10.1093/jac/dkp507
  8. Hoenigl M, Salzer HJ, Raggam RB, Valentin T, Rohn A, Woelfler A, et al. 2012. Impact of galactomannan testing on the prevalence of invasive aspergillosis in patients with hematological malignancies. Med. Mycol. 50: 266-269. https://doi.org/10.3109/13693786.2011.603102
  9. Leeflang MM, Debets-Ossenkopp YJ, Visser CE, Scholten RJ, Hooft L, Bijlmer HA, et al. 2008. Galactomannan detection for invasive aspergillosis in immunocompromised patients. Cochrane Database Syst. Rev. 8: CD007394.
  10. Leenders AC, de Marie S, ten Kate MT, Bakker-Woudenberg IA, Verbrugh HA. 1996. Liposomal amphotericin B (AmBisome) reduces dissemination of infection as compared with amphotericin B deoxycholate (Fungizone) in a rate model of pulmonary aspergillosis. J. Antimicrob. Chemother. 38: 215-225. https://doi.org/10.1093/jac/38.2.215
  11. Lengerova M, Kocmanova I, Racil Z, Hrncirova K, Pospisilova S, Mayer J, et al. 2012. Detection and measurement of fungal burden in a guinea pig model of invasive pulmonary aspergillosis by novel quantitative nested real-time PCR compared with galactomannan and (1,3)-beta-D-glucan detection. J. Clin. Microbiol. 50: 602-608. https://doi.org/10.1128/JCM.05356-11
  12. Liu J, Shi Y, He D, Liu P, Zhang Y, Xu L, Wang L. 2009. Development of real-time PCR method to detect Aspergillus fumigates. Biotechnology 19: 34-36. [Article in Chinese]
  13. Loffler J, Hebart H, Schumacher U, Reitze H, Einsele H. 1997. Comparison of different methods for extraction of DNA of fungal pathogens from cultures and blood. J. Clin. Microbiol. 35: 3311-3312.
  14. Luong ML, Clancy CJ, Vadnerkar A, Kwak EJ, Silveira FP, Wissel MC, et al. 2011. Comparison of an Aspergillus realtime polymerase chain reaction assay with galactomannan testing of bronchoalveolar lavage fluid for the diagnosis of invasive pulmonary aspergillosis in lung transplant recipients. Clin. Infect. Dis. 52: 1218-1226. https://doi.org/10.1093/cid/cir185
  15. McCulloch E, Ramage G, Rajendran R, Lappin DF, Jones B, Warn P, et al. 2012. Antifungal treatment affects the laboratory diagnosis of invasive aspergillosis. J. Clin. Pathol. 65: 83-86. https://doi.org/10.1136/jcp.2011.090464
  16. Mennink-Kersten MA, Donnelly JP, Verweij PE. 2004. Detection of circulating galactomannan for the diagnosis and management of invasive aspergillosis. Lancet Infect. Dis. 4: 349-357. https://doi.org/10.1016/S1473-3099(04)01045-X
  17. Millon L, Grenouillet F, Crouzet J, Larosa F, Loewert S, Bellanger AP, et al. 2010. False-positive Aspergillus real-time PCR assay due to a nutritional supplement in a bone marrow transplant recipient with GVH disease. Med. Mycol. 48: 661-664. https://doi.org/10.3109/13693780903451836
  18. Ruping MJ, Vehreschild JJ, Groll A, Lass-Florl C, Ostermann H, Ruhnke M, Cornely OA. 2011. Current issues in the clinical management of invasive aspergillosis - the AGIHO, DMykG, OGMM and PEG web-based survey and expert consensus conference 2009. Mycoses 54: e557-e568. https://doi.org/10.1111/j.1439-0507.2010.01989.x
  19. Scotter JM, Chambers ST. 2005. Comparison of galactomannan detection, PCR-enzyme-linked immunosorbent assay, and real-time PCR for diagnosis of invasive aspergillosis in a neutropenic rat model and effect of caspofungin acetate. Clin. Diagn. Lab. Immunol. 12: 1322-1327.
  20. Suarez F, Lortholary O, Buland S, Rubio MT, Ghez D, Mahe V, et al. 2008. Detection of circulating Aspergillus fumigatus DNA by real-time PCR assay of large serum volumes improves early diagnosis of invasive aspergillosis in highrisk adult patients under hematologic surveillance. J. Clin. Microbiol. 46: 3772-3777. https://doi.org/10.1128/JCM.01086-08
  21. Torelli R, Sanguinetti M, Moody A, Pagano L, Caira M, De Carolis E, et al. 2011. Diagnosis of invasive aspergillosis by a commercial real-time PCR assay for Aspergillus DNA in bronchoalveolar lavage fluid samples from high-risk patients compared to a galactomannan enzyme immunoassay. J. Clin. Microbiol. 49: 4273-4278. https://doi.org/10.1128/JCM.05026-11
  22. von Eiff M, Roos N, Schulten R, Hesse M, Zuhlsdorf M, van de Loo J. 1995. Pulmonary aspergillosis: early diagnosis improves survival. Respiration 62: 341-347. https://doi.org/10.1159/000196477
  23. Walsh TJ, Shoham S, Petraitiene R, Sein T, Schaufele R, Kelaher A, et al. 2004. Detection of galactomannan antigenemia in patients receiving piperacillin-tazobactam and correlations between in vitro, in vivo, and clinical properties of the drugantigen interaction. J. Clin. Microbiol. 42: 4744-4748. https://doi.org/10.1128/JCM.42.10.4744-4748.2004
  24. Walsh TJ, Wissel MC, Grantham KJ, Petraitiene R, Petraitis V, Kasai M, et al. 2011. Molecular detection and speciesspecific identification of medically important Aspergillus species by real-time PCR in experimental invasive pulmonary aspergillosis. J. Clin. Microbiol. 49: 4150-4157. https://doi.org/10.1128/JCM.00570-11
  25. White PL, Perry MD, Moody A, Follett SA, Morgan G, Barnes RA. 2011. Evaluation of analytical and preliminary clinical performance of Myconostica MycAssay Aspergillus when testing serum specimens for diagnosis of invasive aspergillosis. J. Clin. Microbiol. 49: 2169-2174. https://doi.org/10.1128/JCM.00101-11
  26. Yeo SF, Wong B. 2002. Current status of nonculture methods for diagnosis of invasive fungal infection. Clin. Microbiol. Rev. 15: 465-484. https://doi.org/10.1128/CMR.15.3.465-484.2002
  27. Zhang CR, Tang YC, Kawakami K, Zhang TT, Zhang KX, Zhu JX. 2004. An experimental study of the therapeutic effect of interleukin-2 and interleukin-12 with and without amphotericin B on pulmonary fungal infection. Zhonghua Jie He He Hu Xi Za Zhi 27: 234-236. [in Chinese]
  28. Zhao Y, Park S, Warn P, Shrief R, Harrison E, Perlin DS. 2010. Detection of Aspergillus fumigatus in a rat model of invasive pulmonary aspergillosis by real-time nucleic acid sequence-based amplification. J. Clin. Microbiol. 48: 1378-1383. https://doi.org/10.1128/JCM.02214-09

피인용 문헌

  1. CNTF protects neurons from hypoxic injury through the activation of STAT3pTyr705 vol.38, pp.6, 2014, https://doi.org/10.3892/ijmm.2016.2769
  2. Circulating Aspergillus fumigatus DNA Is Quantitatively Correlated to Galactomannan in Serum vol.8, pp.None, 2017, https://doi.org/10.3389/fmicb.2017.02040
  3. Effects of Aspergillus fumigatus on glucocorticoid receptor and β2-adrenergic receptor expression in a rat model of asthma vol.43, pp.4, 2014, https://doi.org/10.1080/01902148.2017.1339142
  4. Investigating the Presence of Aspergillus fumigatus and A. flavus Using Galactomannan Enzyme Assay and TaqMan Real-Time PCR Technique vol.11, pp.2, 2018, https://doi.org/10.5812/jjm.13670