期刊论文详细信息
BMC Infectious Diseases
Evaluation of Entamoeba histolytica recombinant phosphoglucomutase protein for serodiagnosis of amoebic liver abscess
Lim Boon Huat4  Alfonso Olivos-Garcia3  Zeehaida Mohamed2  Foo Phiaw Chong4  See Too Wei Cun4  Rahmah Noordin1  Wong Weng Kin4  Tan Zi Ning4 
[1] Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, 11800, Malaysia;Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, 16150, Malaysia;Departamento de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México, México D.F, 04510, México;School of Health Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, 16150, Malaysia
关键词: rPGM-ELISA;    Diagnosis;    Phosphoglucomutase;    Western blot;    2-DE;    Amoebic liver abscess;    Entamoeba histolytica;   
Others  :  1149253
DOI  :  10.1186/1471-2334-13-144
 received in 2012-12-25, accepted in 2013-03-13,  发布年份 2013
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【 摘 要 】

Background

Amoebic liver abscess (ALA) is the most frequent clinical presentation of extra-intestinal amoebiasis. The diagnosis of ALA is typically based on the developing clinical symptoms, characteristic changes on radiological imaging and serology. Numerous serological tests have been introduced for the diagnosis of ALA, either detecting circulating amoebic antigens or antibodies. However those tests show some pitfalls in their efficacy and/or the preparation of the tests are costly and tedious. The commercial IHA kit that used crude antigen was reported to be useful in diagnosis of ALA, however high antibody background in endemic areas may cause problems in its interpretation. Thus, discovery of well-defined antigen(s) is urgently needed to improve the weaknesses of current serodiagnostic tests.

Methods

Crude antigen of E. histolytica was analysed by 2-DE and Western blot to identify a protein of diagnostic potential for ALA. The corresponding gene of the antigenic protein was then cloned, expressed and the purified recombinant protein was subsequently evaluated for serodiagnosis of ALA in an indirect ELISA format.

Results

Analysis of crude antigen showed that phosphoglucomutase (PGM) has the diagnostic potential. Recombinant PGM (rPGM) showed 79.17% (19/24) sensitivity and 86.67% (195/225) specificity in diagnosis of ALA based on the COV of mean +1SD. There was no significant difference between rPGM-ELISA and IHA diagnostic kit in the diagnosis of ALA in terms of sensitivity and specificity at p-value < 0.05.

Conclusion

In conclusion, rPGM-ELISA is found to be useful for serodiagnosis of ALA. Future studies will determine whether rPGM-ELISA also detects antibodies produced in amoebic dysentery and asymptomatic cases.

【 授权许可】

   
2013 Ning et al.; licensee BioMed Central Ltd.

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【 参考文献 】
  • [1]Pitt HA: Surgical management of hepatic abscesses. World J Surg 1990, 14:498-504.
  • [2]Mathur S, Gehlot RS, Mohta A, Bhargava N: Clinical profile of amoebic liver abscess. JIACM 2002, 3:367-373.
  • [3]Sharma MP, Ahuja V: Amoebic liver abscess. JIACM 2003, 4:107-111.
  • [4]Wells CD, Arguedas M: Amebic liver abscess. South Med J 2004, 97:673-682.
  • [5]Knappik M, Borner U, Jelinek T: Sensitivity and specificity of a new commercial enzyme-linked immunoassay kit for detecting Entamoeba histolytica IgG antibodies in serum samples. Eur J Clin Microbiol Infect Dis 2005, 24:701-703.
  • [6]Robert R, Mahaza C, Bernard C, Buffard C, Senet JM: Evaluation of a new bicolored latex agglutination test for immunological diagnosis of hepatic amoebiasis. J Clin Microbiol 1990, 28:1422-1424.
  • [7]Wong WK, Tan ZN, Othman N, Lim BH, Mohamed Z, Olivos Garcia A, Noordin R: Analysis of Entamoeba histolytica excretory-secretory antigen and identification of a new potential diagnostic marker. Clin Vaccine Immunol 2011, 18:1913-1917.
  • [8]Yang J, Kennedy MT: Evaluation of enzyme-linked immunosorbent assay for the serodiagnosis of amebiasis. J Clin Microbiol 1979, 10:778-785.
  • [9]Lotter H, Mannweiler E, Schreiber M, Tannich E: Sensitive and specific serodiagnosis of invasive amebiasis by using a recombinant surface protein of pathogenic Entamoeba histolytica. J Clin Microbiol 1992, 30:3163-3167.
  • [10]del Carmen S-GM, Velazquez-Rojas M, Salgado-Rosas H, Torres-Rasgado E, Perez-Fuentes R, Martinez-Munguia J, Talamas-Rohana P: Seroprevalence of anti-entamoeba histolytica antibodies by IHA and ELISA assays in blood donors from Puebla. Mexico. Arch Med Res 2000, 31:S53-S54.
  • [11]Kraoul L, Adjmi H, Lavarde V, Pays JF, Tourte-Schaefer C, Hennequin C: Evaluation of a rapid enzyme immunoassay for diagnosis of hepatic amoebiasis. J Clin Microbiol 1997, 35:1530-1532.
  • [12]Zeehaida M, Bachok N, Hasan H: Analysis of indirect hemagglutination assay results among patients with amoebic liver abscess. Int Med J 2009, 16:195-199.
  • [13]Stanley SL Jr: Progress towards development of a vaccine for amebiasis. Clin Microbiol Rev 1997, 10:637-649.
  • [14]Myung K, Burch D, Jackson TF, Stanley SL Jr: Serodiagnosis of invasive amebiasis using a recombinant Entamoeba histolytica antigen-based ELISA. Arch Med Res 1992, 23:285-288.
  • [15]Ravdin JI, Jackson TF, Petri WA Jr, Murphy CF, Ungar BL, Gathiram V, Skilogiannis J, Simjee AE: Association of serum antibodies to adherence lectin with invasive amebiasis and asymptomatic infection with pathogenic Entamoeba histolytica. J Infect Dis 1990, 162:768-772.
  • [16]Zhang Y, Li E, Jackson TF, Zhang T, Gathiram V, Stanley SL Jr: Use of a recombinant 170-kilodalton surface antigen of Entamoeba histolytica for serodiagnosis of amebiasis and identification of immunodominant domains of the native molecule. J Clin Microbiol 1992, 30:2788-2792.
  • [17]Diamond LS, Harlow DR, Cunnick CC: A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 1978, 72:431-432.
  • [18]Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976, 72:248-254.
  • [19]Reen DJ: Basic Protein and Peptide Protocols: CHAPTER 47 Enzyme-Linked immunosorbent Assay (ELISA). Totowa, NJ: Humana Press Inc.; 1994.
  • [20]Keeley KJ, Schmaman A, Scott A: Definitive diagnosis of amoebic liver abscess: value of liver biopsy. Br Med J 1962, 1:375-376.
  • [21]Tsutsumi V, Mena-Lopez R, Anaya-Velazquez F, Martinez-Palomo A: Cellular bases of experimental amebic liver abscess formation. Am J Pathol 1984, 117:81-91.
  • [22]Fotedar R, Stark D, Beebe N, Marriott D, Ellis J, Harkness J: Laboratory diagnostic techniques for Entamoeba species. Clin Microbiol Rev 2007, 20:511-532. table of contents
  • [23]Haque R, Mollah NU, Ali IK, Alam K, Eubanks A, Lyerly D, Petri WA Jr: Diagnosis of amebic liver abscess and intestinal infection with the TechLab Entamoeba histolytica II antigen detection and antibody tests. J Clin Microbiol 2000, 38:3235-3239.
  • [24]Parija SC, Khairnar K: Detection of excretory Entamoeba histolytica DNA in the urine, and detection of E. histolytica DNA and lectin antigen in the liver abscess pus for the diagnosis of amoebic liver abscess. BMC Microbiol 2007, 7:41. BioMed Central Full Text
  • [25]Zeehaida M, Wan Nor Amilah WA, Amry AR, Hassan S, Sarimah A, Rahmah N: A study on the usefulness of Techlab Entamoeba histolytica II antigen detection ELISA in the diagnosis of amoebic liver abscess (ALA) at Hospital Universiti Sains Malaysia (HUSM), Kelantan, Malaysia. Trop Biomed 2008, 25:209-216.
  • [26]Valenzuela O, Ramos F, Morán P, González E, Valadez A, Gómez A, Melendro EI, Ramiro M, Muñoz O, Ximénez C: Persistence of secretory antiamoebic antibodies in patients with past invasive intestinal or hepatic amoebiasis. Parasitol Res 2001, 87:849-852.
  • [27]Haghighi A, Rezaeian M: Detection of serum antibody to Entameba histolytica in various population samples of amebic infection using an enzyme-linked immunosorbent assay. Parasitol Res 2005, 97:209-212.
  • [28]Shenai BR, Komalam BL, Arvind AS, Krishnaswamy PR, Rao PV: Recombinant antigen-based avidin-biotin microtiter enzyme-linked immunosorbent assay for serodiagnosis of invasive amebiasis. J Clin Microbiol 1996, 34:828-833.
  • [29]Tachibana H, Cheng XJ, Masuda G, Horiki N, Takeuchi T: Evaluation of recombinant fragments of Entamoeba histolytica Gal/GalNAc lectin intermediate subunit for serodiagnosis of amebiasis. J Clin Microbiol 2004, 42:1069-1074.
  • [30]Pal S, Sengupta K, Manna B, Sarkar S, Bhattacharya S, Das P: Comparative evaluation of somatic & excretory-secretory antigens of Entamoeba histolytica in serodiagnosis of human amoebiasis by ELISA. Indian J Med Res 1996, 104:152-156.
  • [31]Sathar MA, Bredenkamp BL, Gathiram V, Simjee AE, Jackson TF: Detection of Entamoeba histolytica immunoglobulins G and M to plasma membrane antigen by enzyme-linked immunosorbent assay. J Clin Microbiol 1990, 28:332-335.
  • [32]Zhou M, Conrads TP, Veenstra TD: Proteomics approaches to biomarker detection. Brief Funct Genomic Proteomic 2005, 4:69-75.
  • [33]Ortner S, Binder M, Scheiner O, Wiedermann G, Duchene M: Molecular and biochemical characterization of phosphoglucomutases from Entamoeba histolytica and Entamoeba dispar. Mol Biochem Parasitol 1997, 90:121-129.
  • [34]Flores BM, Reed SL, Ravdin JI, Torian BE: Serologic reactivity to purified recombinant and native 29-kilodalton peripheral membrane protein of pathogenic Entamoeba histolytica. J Clin Microbiol 1993, 31:1403-1407.
  • [35]Kaur U, Sharma AK, Sharma M, Vohra H: Distribution of Entamoeba histolytica Gal/GalNAc lectin-specific antibody response in an endemic area. Scand J Immunol 2004, 60:524-528.
  • [36]Sanuki J, Nakano K, Tokoro M, Nozaki T, Okuzawa E, Kobayashi S, Asai T: Purification and identification of major soluble 40-kDa antigenic protein from Entamoeba histolytica: its application for serodiagnosis of asymptomatic amebiasis. Parasitol Int 2001, 50:73-80.
  • [37]Abd-Alla MD, Jackson TF, Reddy S, Ravdin JI: Diagnosis of invasive amebiasis by enzyme-linked immunosorbent assay of saliva to detect amebic lectin antigen and anti-lectin immunoglobulin G antibodies. J Clin Microbiol 2000, 38:2344-2347.
  • [38]Lee J, Park SJ, Yong TS: Serodiagnosis of amoebiasis using a recombinant protein fragment of the 29 kDa surface antigen of Entamoeba histolytica. Int J Parasitol 2000, 30:1487-1491.
  • [39]Stanley SL Jr, Jackson TF, Reed SL, Calderon J, Kunz-Jenkins C, Gathiram V, Li E: Serodiagnosis of invasive amebiasis using a recombinant Entamoeba histolytica protein. JAMA 1991, 266:1984-1986.
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