Comparative Evaluation of Serological, ELISA and Molecular Tests in Diagnosis of Brucella abortus in Human Serum
,
Abstract
Background: Brucellosis is one of the most prevalent and common diseases between humans and animals.Prompt diagnosis and timely treatment of this disease can prevent many complications. In this regard, this study aims to comparatively evaluate ELISA, PCR and serological methods to identify Brucella abortus.
Methods: In this study, the serum of 100 patients referred to Tonekabon private laboratory from July 2020 to January 2021 was examined by PCR, ELISA and Wright, Coombs Wright, and 2ME methods for the detection of Brucella abortus.
Results: In this study, the mean age of the sample was 43.3 ± 18.2 of which 21% were infected with Brucella abortus according to the above serological methods. According to ELISA test, 22% of the samples were IgM, 6% of the samples were IgG and 16% were PCR positive. Kappa agreement coefficient in Wright and Coombs Wright test and 2me were significant (P <0.001). Serological diagnostic indices and ELISA sensitivity were 68.75% and 68.75%, respectively. The lowest prediction rate of Brucella abortus among diagnostic methods was related to Elisa (IgM). Based on Fisher's exact test, there was no significant relationship between the percentage of Brucella abortus positive PCR cases and age, sex, previous history of infection with Brucella, fever, body aches and dairy consumption.
Conclusion: Based on the results of our study, the accuracy of all methods is comparative and the lowest accuracy is related to Elisa (IgM) which has a lower level of predictability than other methods. The highest level of prediction belonged to Wright and 2me tests.
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2. Akbarzadeh E, Noroosi J, Piranfar V, et al. Simultaneous identification and comparison of pathogenic Brucella species in human serum and blood samples by means of multiplex PCR and confirmation of the results by PCR-RFLP. SJKU 2015; 20(1):10-17.
3. Zeinali M. Iranin national of guideline for brucellosis control. Ministery of health and medical education Health deputy center for disease control zoonoses office. 2012:17-18.
4. Najafi N, Ghassemian R, Davoody AR, et al. An unusual complication of a common endemic disease: clinical and laboratory aspects of patients with Brucella epididymoorchitis in the north of Iran. BMC Research Notes 2011; 4(1):286.
5. Mirnejad R, Mohamadi M, Piranfar V, et al. A duplex PCR for rapid and simultaneous detection of Brucella spp. in human blood samples. Asian Pac J Trop Med 2013; 6(6):453-56.
6. Mirnejad R, Jazi FM, Mostafaei S, et al. Epidemiology of brucellosis in Iran: A comprehensive systematic review and metaanalysis study. Microb Pathog 2017; 109:239-247.
7. Saboura S, Arzanloua M, Jeddib F, et al. Evaluating the efficiency of Taq Man real-time ocr and serological methods in the detectiin of Brucella spp. in clinical speciemens. J Microbiol Methods 2020; 175:105982
8. Ghorbani A, Khorasgani MR, Zarkesh Esfahani H, et al. Comparison of serology, calture, and PCR for detection of brucellosis in staughtered camels in Iran. Camp clin pathol 2013; 22:913-917.
9. Khademi, F, Yousefi-Avarvand A, Sahebkar A, et al. Drug resistance of clinical and environmental isolates of Brucella species in Iran: a meta-analysis. Rev Med Microbiol 2018; 29:166-72.
10. Khan AU, Melzer F, El-Soally S, et al. Serological and molecular identification of Brucella spp. in pigs from Cairo and Giza Governorates, Egypt. Pathogens 2019; 8, 248.
11. Bhat I, Mashooq M, Kumar D, et al. Development of probebased real-time loop-mediated isothermal amplification for detection of Brucella. J Appl Microbiol 2019; 126:1332-9.
12. Roushan MRH, Amiri MJS, Laly A, et al. Follow-up standard agglutination and 2-mercaptoethanol tests in 175 clinically cured cases of human brucellosis. Int J Infect Dis 2010; 14:e250-e253.
13. Gad EL, Rab MO. Evaluation of Brucella enzyme immunoassay test (ELISA) in compatison with bacteriological culture and agglitination. J Infect Dev 1998; 36(2):197-201.
14. Yagupsky P, Morata P, Colmenero JD. Laboratory diagnosis of human brucellosis Pablo. Clin Microbiol Rev 2020; 33:1-54 .
15. Zeybek H, Acikgoz ZC, Dal T, et al. Optimization and validation of a real-time polymerase chain reaction protocol for the diagnosis of human brucellosis. Folia Microbiol 2020; 65(2):353-61.
16. Lazutkin A, korem M, Weinberger J, et al. Otolaryngology/head and neck region manifestations of Brucella. The Laryngoscope 2018; 128(9):2056-9.
17. Kiambi SG, Fèvre EM, Omolo J, et al. Risk factors for acute human brucellosis in Ijara, north-eastern Kenya. PLoS Negl Trop Dis 2020; 14:e0008108.
18. Bodenham FR, Lukambagire AB, Ashfor RT, et al. Prevalence and speciation of acute brucellosis in febrile patients from a pastoralist community of Tanzania. Sci Rep 2020; 10:1-10.
19. Haghdoost M, Ansari L, Waysee Osquee H. Comparison of Brucella capture and Coombs wright and wright in the diagnosis of Brucellosis. J Res clin Med 2021; 9(1):15-15
20. Arabi MR, Karami M, Keramat F, et al. Serological and molecular investigation of human brucellosis in participants of Famenin brucellosis cohort study, Hamadan, Iran. Iran J Microbio 2021; 13(3):319-324.
21. Shalini T, Jabir S. Singh SR, et al.Quantitative polymerase chain reaction based quantification of Brucella DNA in serum of pre-and post-therapeutic occupationally exposed infected human population. J Infec Pub Health 2017; 11:514-520.
22. Sabour S, Arzanlou M, Jeddi F, et al. Evaluting the efficiency of Taq Man real-time PCR and serological methods in the detection of Brucella spp.in clinical specimens collected from suspected patients in Ardabil, Iran. J Microbiol Methods 2020; 175,115-105982.
23. Arbab S, Shahzad A, Akhter S, et al. Acute febrile ilness caused by Brucella abortus infectiin in human in pakistan. IJERPH 2019; 25(1):250-9.
24. Piranfar V, Sharif M, Hashemi M, et al. Detection and discrimination of two Brucella species by moltiplex real -time PCR and high-resolution meta analysis curve from human blood abd comparison of results using RFLP. Iran J Basic Med sci 2015; 18(9):909-14.
25. Mohseni KH, Mirnejad R, Piranfar V, et al. Acomparative evalution of Elisa, PCR and serum agglutination tests for diagnosis of Brucella using human serum. Iran J Pathol 2017; 12(4):371.
26. Hajia M, Fallah F, Angoti G, et al. Comparison of methods for diagnosing brucellosis. Lab Med 2013; 44(1):20-23.
27. Nannan W, Wei W, Fengzhe C, et al. Elisa is superior to bacterial calture and agglutination test in tge diagnosis of brucellosis in an endemic areain china. BMC infect dis 2020; 11(20):1-7.
28. Najafi N, Davoodi L, Fazki M, et al. Diagnostic value of Elisa versus wright in human brucellosis with positive pcr. J Mazandaran Univ Med Sci 2013; 2:263-268.
29. Neha VAK, Kumar A, Ahmed I. Comparative efficacy of serological diagnostic methods and evalution of polymerase chain reaction for diagnosis of bovine brucellosis . Iran J Vet Res 2017; 21:279-281.
30. Kumar A, Gupta VK, Verma AK, et al. Seroprevalence and risk factors associated with bovine brucellosis in western Uttar Pradesh, India. Ind J Anim Sci 2016; 86:131- 135.
31. Neha AW, Verma AK, Jain U, et al. Brucellosis in organized dairy farm: an investigation. Asian J Anim Sci 2014; 8:29-33.
32. Verma AK, Dhama K, Chakraborty S, et al. Strategies for combating and eradicating important infectious diseases of animals with particular reference to India: present and future perspectives. Asian J Anim Vet Adv 2014; 9:77-106.
33. Tabasi M, Eybpoosh S, Bouzari S. Development of and indirect Elisa based on whole cell Brucella abortus s99 lysates for detection of IgM anti -Brucella antibodies in human serum. Comp Immunol Microbiol Infect 2019; 63:87-93.
34. Pakzad I, Bahmani S, Ghafouryan S, et al. Diagnosis of human brucellosis by pcr using 17/112 and 16 sr RNA genes compared with common serological tests. J Arak Uni Med Sci 2012; 14;112-120.
35. Rajeswari Sh, Triveni K, Krithig N. Serological and molecular analysis for brucellosis in selected swine herds from Southern India. J Infect Public Health 2019; 12: 247-51.
36. Mitka S, Anetakis C, Souliou E, et al. Evaluation of different PCR assays for early detection of acute and relapsing brucellosis in humans in comparsion with conventional methods. J Clin Microbiol 2007; 45(4):1211-8.
37. Abdelbaset EA, Abushaba M, Hamed M, et al. Sero-diagnosis of brucellosis in sheep and humans in Assiut and El-Minya governorates, Egypt. Int J Vet Sci 2018; 6;63-67.
38. Agin M, Kayar Y. Demographic, laboratory, and clinical comparison of pediatric Brucella cases with and without liver Involvement. Cureus 2020; 21:147-52.
39. Shahzad A, Usama S, Rizwan M, et al. Serosurvey and risk factors associated with Brucella infection in high risk occupations from district Lahore and Kasur of Punjab, Pakistan. Pathogens 2021; 61:152-9.
40. Munyua P, Osoro E, Hunsberger E, et al. High incidence of human brucellosis in a rural pastoralist community in kenya. PLoS Negl Trop Dis 2015; 15(2):35-9.
2. Akbarzadeh E, Noroosi J, Piranfar V, et al. Simultaneous identification and comparison of pathogenic Brucella species in human serum and blood samples by means of multiplex PCR and confirmation of the results by PCR-RFLP. SJKU 2015; 20(1):10-17.
3. Zeinali M. Iranin national of guideline for brucellosis control. Ministery of health and medical education Health deputy center for disease control zoonoses office. 2012:17-18.
4. Najafi N, Ghassemian R, Davoody AR, et al. An unusual complication of a common endemic disease: clinical and laboratory aspects of patients with Brucella epididymoorchitis in the north of Iran. BMC Research Notes 2011; 4(1):286.
5. Mirnejad R, Mohamadi M, Piranfar V, et al. A duplex PCR for rapid and simultaneous detection of Brucella spp. in human blood samples. Asian Pac J Trop Med 2013; 6(6):453-56.
6. Mirnejad R, Jazi FM, Mostafaei S, et al. Epidemiology of brucellosis in Iran: A comprehensive systematic review and metaanalysis study. Microb Pathog 2017; 109:239-247.
7. Saboura S, Arzanloua M, Jeddib F, et al. Evaluating the efficiency of Taq Man real-time ocr and serological methods in the detectiin of Brucella spp. in clinical speciemens. J Microbiol Methods 2020; 175:105982
8. Ghorbani A, Khorasgani MR, Zarkesh Esfahani H, et al. Comparison of serology, calture, and PCR for detection of brucellosis in staughtered camels in Iran. Camp clin pathol 2013; 22:913-917.
9. Khademi, F, Yousefi-Avarvand A, Sahebkar A, et al. Drug resistance of clinical and environmental isolates of Brucella species in Iran: a meta-analysis. Rev Med Microbiol 2018; 29:166-72.
10. Khan AU, Melzer F, El-Soally S, et al. Serological and molecular identification of Brucella spp. in pigs from Cairo and Giza Governorates, Egypt. Pathogens 2019; 8, 248.
11. Bhat I, Mashooq M, Kumar D, et al. Development of probebased real-time loop-mediated isothermal amplification for detection of Brucella. J Appl Microbiol 2019; 126:1332-9.
12. Roushan MRH, Amiri MJS, Laly A, et al. Follow-up standard agglutination and 2-mercaptoethanol tests in 175 clinically cured cases of human brucellosis. Int J Infect Dis 2010; 14:e250-e253.
13. Gad EL, Rab MO. Evaluation of Brucella enzyme immunoassay test (ELISA) in compatison with bacteriological culture and agglitination. J Infect Dev 1998; 36(2):197-201.
14. Yagupsky P, Morata P, Colmenero JD. Laboratory diagnosis of human brucellosis Pablo. Clin Microbiol Rev 2020; 33:1-54 .
15. Zeybek H, Acikgoz ZC, Dal T, et al. Optimization and validation of a real-time polymerase chain reaction protocol for the diagnosis of human brucellosis. Folia Microbiol 2020; 65(2):353-61.
16. Lazutkin A, korem M, Weinberger J, et al. Otolaryngology/head and neck region manifestations of Brucella. The Laryngoscope 2018; 128(9):2056-9.
17. Kiambi SG, Fèvre EM, Omolo J, et al. Risk factors for acute human brucellosis in Ijara, north-eastern Kenya. PLoS Negl Trop Dis 2020; 14:e0008108.
18. Bodenham FR, Lukambagire AB, Ashfor RT, et al. Prevalence and speciation of acute brucellosis in febrile patients from a pastoralist community of Tanzania. Sci Rep 2020; 10:1-10.
19. Haghdoost M, Ansari L, Waysee Osquee H. Comparison of Brucella capture and Coombs wright and wright in the diagnosis of Brucellosis. J Res clin Med 2021; 9(1):15-15
20. Arabi MR, Karami M, Keramat F, et al. Serological and molecular investigation of human brucellosis in participants of Famenin brucellosis cohort study, Hamadan, Iran. Iran J Microbio 2021; 13(3):319-324.
21. Shalini T, Jabir S. Singh SR, et al.Quantitative polymerase chain reaction based quantification of Brucella DNA in serum of pre-and post-therapeutic occupationally exposed infected human population. J Infec Pub Health 2017; 11:514-520.
22. Sabour S, Arzanlou M, Jeddi F, et al. Evaluting the efficiency of Taq Man real-time PCR and serological methods in the detection of Brucella spp.in clinical specimens collected from suspected patients in Ardabil, Iran. J Microbiol Methods 2020; 175,115-105982.
23. Arbab S, Shahzad A, Akhter S, et al. Acute febrile ilness caused by Brucella abortus infectiin in human in pakistan. IJERPH 2019; 25(1):250-9.
24. Piranfar V, Sharif M, Hashemi M, et al. Detection and discrimination of two Brucella species by moltiplex real -time PCR and high-resolution meta analysis curve from human blood abd comparison of results using RFLP. Iran J Basic Med sci 2015; 18(9):909-14.
25. Mohseni KH, Mirnejad R, Piranfar V, et al. Acomparative evalution of Elisa, PCR and serum agglutination tests for diagnosis of Brucella using human serum. Iran J Pathol 2017; 12(4):371.
26. Hajia M, Fallah F, Angoti G, et al. Comparison of methods for diagnosing brucellosis. Lab Med 2013; 44(1):20-23.
27. Nannan W, Wei W, Fengzhe C, et al. Elisa is superior to bacterial calture and agglutination test in tge diagnosis of brucellosis in an endemic areain china. BMC infect dis 2020; 11(20):1-7.
28. Najafi N, Davoodi L, Fazki M, et al. Diagnostic value of Elisa versus wright in human brucellosis with positive pcr. J Mazandaran Univ Med Sci 2013; 2:263-268.
29. Neha VAK, Kumar A, Ahmed I. Comparative efficacy of serological diagnostic methods and evalution of polymerase chain reaction for diagnosis of bovine brucellosis . Iran J Vet Res 2017; 21:279-281.
30. Kumar A, Gupta VK, Verma AK, et al. Seroprevalence and risk factors associated with bovine brucellosis in western Uttar Pradesh, India. Ind J Anim Sci 2016; 86:131- 135.
31. Neha AW, Verma AK, Jain U, et al. Brucellosis in organized dairy farm: an investigation. Asian J Anim Sci 2014; 8:29-33.
32. Verma AK, Dhama K, Chakraborty S, et al. Strategies for combating and eradicating important infectious diseases of animals with particular reference to India: present and future perspectives. Asian J Anim Vet Adv 2014; 9:77-106.
33. Tabasi M, Eybpoosh S, Bouzari S. Development of and indirect Elisa based on whole cell Brucella abortus s99 lysates for detection of IgM anti -Brucella antibodies in human serum. Comp Immunol Microbiol Infect 2019; 63:87-93.
34. Pakzad I, Bahmani S, Ghafouryan S, et al. Diagnosis of human brucellosis by pcr using 17/112 and 16 sr RNA genes compared with common serological tests. J Arak Uni Med Sci 2012; 14;112-120.
35. Rajeswari Sh, Triveni K, Krithig N. Serological and molecular analysis for brucellosis in selected swine herds from Southern India. J Infect Public Health 2019; 12: 247-51.
36. Mitka S, Anetakis C, Souliou E, et al. Evaluation of different PCR assays for early detection of acute and relapsing brucellosis in humans in comparsion with conventional methods. J Clin Microbiol 2007; 45(4):1211-8.
37. Abdelbaset EA, Abushaba M, Hamed M, et al. Sero-diagnosis of brucellosis in sheep and humans in Assiut and El-Minya governorates, Egypt. Int J Vet Sci 2018; 6;63-67.
38. Agin M, Kayar Y. Demographic, laboratory, and clinical comparison of pediatric Brucella cases with and without liver Involvement. Cureus 2020; 21:147-52.
39. Shahzad A, Usama S, Rizwan M, et al. Serosurvey and risk factors associated with Brucella infection in high risk occupations from district Lahore and Kasur of Punjab, Pakistan. Pathogens 2021; 61:152-9.
40. Munyua P, Osoro E, Hunsberger E, et al. High incidence of human brucellosis in a rural pastoralist community in kenya. PLoS Negl Trop Dis 2015; 15(2):35-9.
| Files | ||
| Issue | Vol 11 No 5-6 (2023) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/jmb.v11i5-6.14361 | |
| Keywords | ||
| Brucella abortus ELISA Molecular Diagnosis Serological. | ||
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How to Cite
1.
Ghanbari F, Fakhar H, Alamian S, Shaghaghi B. Comparative Evaluation of Serological, ELISA and Molecular Tests in Diagnosis of Brucella abortus in Human Serum. J Med Bacteriol. 2023;11(5-6):38-48.
