Evaluation of Complex and Defined Antigens of &lt;i&gt;Mycobacterium tuberculosi&lt;/i&gt;&lt;i&gt;s&lt;/i&gt; in an IgG-Specific ELISA for the Diagnosis of Tuberculosis

Amoudy, Hanady A.; Al-Asmer, Abu B.H.; Abul, Adnan T.; Mustafa, Abu Salim

doi:10.1159/000157434

Cited by 10 publications

(8 citation statements)

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“…Two precipitin bands of identity were observed with the serum of our patient. It was further confirmed by demonstrating elevated levels of anti B. ranarumspecific immunoglobulin M (IgM) and IgG antibodies by a direct immunosorbent assay (enzyme-linked immunosorbent assay [ELISA]) described previously for the detection of antibodies to Mycobacterium tuberculosis (1). By using locally de- rived B. ranarum antigen in the ELISA, only IgG1 and IgG3 titers were selectively elevated in the patient's serum.…”

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confidence: 69%

Basidiobolus ranarum as an Etiologic Agent of Gastrointestinal Zygomycosis

Khoursheed²,

et al. 2001

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Basidiobolus ranarum is a known cause of subcutaneous zygomycosis. Recently, its etiologic role in gastrointestinal infections has been increasingly recognized. While the clinical presentation of the subcutaneous disease is quite characteristic and the disease is easy to diagnose, gastrointestinal basidiobolomycosis poses diagnostic difficulties; its clinical presentation is nonspecific, there are no identifiable risk factors, and all age groups are susceptible. The case of gastrointestinal basidiobolomycosis described in the present report occurred in a 41-year-old Indian male who had a history of repair of a left inguinal hernia 2 years earlier and who is native to the southern part of India, where the subcutaneous form of the disease is indigenous. Diagnosis is based on the isolation of B. ranarum from cultures of urine and demonstration of broad, sparsely septate hyphal elements in histopathologic sections of the colon, with characteristic eosinophilic infiltration and the Splendore-Hoeppli phenomenon. The titers of both immunoglobulin G (IgG) and IgM antibodies to locally produced antigen of the fungus were elevated. The patient failed to respond to 8 weeks of amphotericin B therapy, and the isolate was later found to be resistant to amphotericin B, itraconazole, fluconazole, and flucytosine but susceptible to ketoconazole and miconazole. One other noteworthy feature of the fungus was that the patient's serum showed raised levels of Th2-type cytokines (interleukins 4 and 10) and tumor necrosis factor alpha. The present report underscores the need to consider gastrointestinal basidiobolomycosis in the differential diagnosis of inflammatory bowel diseases and suggests that, perhaps, more time should be invested in developing standardized serologic reagents that can be used as part of a less invasive means of diagnosis of the disease. CASE REPORTA 41-year-old, Indian male was admitted to Mubarak AlKabeer Hospital, Kuwait, in December 1999 with a history of abdominal pain of 20 days' duration. On examination, he was febrile (38.8°C) and his abdomen was tender and distended, particularly at the right hypochondrium and the right iliac fossa, where a palpable, nodular mass (9 by 5 cm) was located. The patient's history was not remarkable, except that he had a history of repair of a left inguinal hernia 2 years earlier. He was nondiabetic and nonneutropenic but had persistently low hemoglobin levels (7.7 to 8.7 g/dl). Ultrasound of the abdomen and pelvis confirmed the presence of a thick mass in the right iliac fossa and showed a marked thickening of the ascending colon and cecum and an increased echogenicity of the renal cortex, suggesting bilateral compromised function. Moreover, his prostate was markedly enlarged. There was fecal impaction in the colon. His blood urea nitrogen, serum creatinine, and uric acid levels were within the normal ranges. Examination of his urine showed many leukocytes and red blood cells, besides elevated levels of protein and bilirubin. On 11 January 2000, a right hemicolectomy was per...

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confidence: 69%

Basidiobolus ranarum as an Etiologic Agent of Gastrointestinal Zygomycosis

Khoursheed²,

et al. 2001

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“…The complex antigens were killed whole cells of M. tuberculosis H37Ra [11] , M. tuberculosis culture filtrate (MT-CF) highly enriched for secreted antigens and purified M. tuberculosis cell walls (MT-CW) [12] . MT-CF and MT-CW were kindly provided by Dr. P.J.…”

Section: Complex and Fractionated Mycobacterial Antigensmentioning

confidence: 99%

Cell-Mediated Immune Responses to Complex and Single Mycobacterial Antigens in Tuberculosis Patients with Diabetes

et al. 2008

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Objective: To evaluate cell-mediated immune (CMI) response in diabetic and non-diabetic tuberculosis (TB) patients and healthy subjects in response to complex, fractionated and single antigens of Mycobacteriumtuberculosis. Material and Methods: Peripheral blood mononuclear cells (PBMC) were obtained from patients suffering from pulmonary TB and type II diabetes (n = 7), pulmonary TB without diabetes (n = 10) and healthy subjects without TB and diabetes (n = 10). PBMC were assessed for CMI responses in antigen-induced proliferation assays in response to complex mycobacterial antigens (whole cells, cell walls and culture filtrate of M. tuberculosis), a battery of naturally purified or recombinant produced secreted (ESAT6, MPT59, MPT64 and MTB38) and cytosolic (MTB10, MTB70, ML10, ML28, ML36, ML65 and MB65) mycobacterial antigens and fractionated culture filtrate proteins (fractions F1–F10) of M. tuberculosis. Results:The majority (>70%) of diabetic and non-diabetic TB patients and healthy subjects responded to the complex antigens of M. tuberculosis. However, among the single antigens, ESAT6 was most frequently recognized by TB patients with and without diabetes, but least recognized by healthy subjects. The secreted antigens MPT59 and MPT64 were recognized by all the groups, whereas the cytosolic antigens were recognized best by healthy subjects. When tested with fractionated secreted proteins present in the culture filtrate of M. tuberculosis, the best responses in both diabetic and non-diabetic TB patients were obtained with fractions containing low-molecular-weight proteins. Conclusions: Diabetic and non-diabetic TB patients respond frequently to secreted low-molecular-weight ESAT6 antigen of M. tuberculosis, indicating that this antigen may be useful in the diagnosis of TB in both the groups.

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“…Several antigens of M. tuberculosis that elicit M. tuberculosisspecific immune responses and/or are involved in protective immunity against TB have been the subject of numerous studies [3][4][5][6][7]. Protein antigens present in the culture filtrate of M. tuberculosis have been studied in great detail mainly due to their ability to induce strong immune responses and to elicit protective immunity in animal models of TB [7][8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Identification of a Novel Protein Antigen Encoded by a Mycobacterium tuberculosis‐Specific RD1 Region Gene

et al. 1999

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A genomic DNA region, designated RD1, that is present in virulent and clinical strains of Mycobacterium tuberculosis and M. bovis, has been shown to be deleted in bacillus Calmette Guérin (BCG). The DNA segments corresponding to three open reading frames (ORFs: ORF‐10, ORF‐14 and ORF‐15) of the RD1 region, that are deleted in BCG strains, were amplified from M. tuberculosis genomic DNA by polymerase chain reaction (PCR), subcloned into pGEX‐4T vector system and expressed in Escherichia coli as fusion proteins with glutathione‐S‐transferase (GST). The recombinant proteins appeared as major cellular proteins in SDS‐PAGE gels at the expected molecular mass. The identity of each fusion protein was confirmed by reactivity with anti‐GST antibodies in Western immunoblots. When pooled human sera from 11 tuberculosis (TB) patients were used as the source of antibodies, only GST‐ORF‐14 fusion protein reacted in Western immunoblots. The protein corresponding to ORF‐14 was then purified to near homogeneity and isolated free of its fusion partner (GST) by treating the purified GST‐ORF‐14 fusion protein with thrombin protease. In Western immunoblots, the purified ORF‐14 protein reacted with antibodies in 26 of 57 human sera (46%) from TB patients while no reactivity was seen with 11 sera from M. bovis BCG‐vaccinated healthy subjects. Interestingly, sera from nine of 15 (60%) long‐term contacts of TB patients also had antibodies reactive to the ORF‐14 protein. These results suggest that the ORF‐14 protein in combination with other immunodominant proteins could be useful in the serodiagnosis of individuals infected with M. tuberculosis.

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Evaluation of Complex and Defined Antigens of <i>Mycobacterium tuberculosi</i><i>s</i> in an IgG-Specific ELISA for the Diagnosis of Tuberculosis

Cited by 10 publications

References 4 publications

Basidiobolus ranarum as an Etiologic Agent of Gastrointestinal Zygomycosis

Basidiobolus ranarum as an Etiologic Agent of Gastrointestinal Zygomycosis

Cell-Mediated Immune Responses to Complex and Single Mycobacterial Antigens in Tuberculosis Patients with Diabetes

Identification of a Novel Protein Antigen Encoded by a Mycobacterium tuberculosis‐Specific RD1 Region Gene

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