Immuno-PCR: a promising ultrasensitive diagnostic method to detect antigens and antibodies

Malou, Nada; Raoult, Didier

doi:10.1016/j.tim.2011.03.004

Cited by 109 publications

(84 citation statements)

References 55 publications

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“…The total plasmid content of each inoculum was confirmed to be as expected (20). Whole-blood samples were collected from the three inoculated mice as well as one noninoculated mouse by submandibular bleed preinoculation and at days 1,3,4,7,9,11,14,16,18, and 21 postinfection. The coagulated blood was spun at 4,000 ϫ g for 9 min to prepare serum.…”

Section: Methodsmentioning

confidence: 95%

Enhanced Detection of Host Response Antibodies to Borrelia burgdorferi Using Immuno-PCR

Halpern¹,

Jain²,

Jewett³

2013

Clin. Vaccine Immunol.

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Lyme disease is the fastest-growing zoonotic disease in North America. Current methods for detection of Borrelia burgdorferi infection are challenged by analysis subjectivity and standardization of antigen source. In the present study, we developed an immuno-PCR (iPCR)-based approach employing recombinant in vivo-expressed B. burgdorferi antigens for objective detection of a host immune response to B. burgdorferi infection. iPCR is a liquid-phase protein detection method that combines the sensitivity of PCR with the specificity and versatility of immunoassay-based protocols. Use of magnetic beads coated with intact spirochetes provided effective antigen presentation and allowed detection of host-generated antibodies in experimentally infected mice at day 11 postinoculation, whereas host-generated antibodies were detected at day 14 by enzyme-linked immunosorbent assay (ELISA) and day 21 by immunoblotting. Furthermore, magnetic beads coated with recombinant B. burgdorferi in vivoexpressed antigen OspC or BmpA demonstrated positive detection of host-generated antibodies in mice at day 7 postinoculation with markedly increased iPCR signals above the background, with the quantification cycle (C q ) value for each sample minus the mean background C q plus 3 standard deviations (⌬C q ) being 4 to 10, whereas ⌬C q was 2.5 for intact spirochete-coated beads. iPCR demonstrated a strong correlation (Spearman rank correlation ‫؍‬ 0.895, P < 0.0001) with a commercial ELISA for detection of host antibodies in human Lyme disease patient sera using the B. burgdorferi VlsE C6 peptide. In addition, iPCR showed potential applicability for direct detection of spirochetes in blood. The results presented here indicate that our iPCR assay has the potential to provide an objective format that can be used for sensitive detection of multiple host response antibodies and isotypes to B. burgdorferi infection.

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Section: Methodsmentioning

confidence: 95%

Enhanced Detection of Host Response Antibodies to Borrelia burgdorferi Using Immuno-PCR

Halpern¹,

Jain²,

Jewett³

2013

Clin. Vaccine Immunol.

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“…To optimize the immuno-PCR conditions, especially to reduce the background noise because of high sensitivity, 13 we adjusted the washing reagents, washing times, blocking reagents, and the concentrations of first and second antibody, and found the following optimized condition. The monoclonal antibody 3A4 specific to PfHRP-2 and PfHRP-3 detection described previously was used as the capture antibody.…”

Section: Methodsmentioning

confidence: 99%

“…12 Immuno-PCR has been applied as a diagnostic test to detect several viral antigens (e.g., antigens of hepatitis B, Rotavirus, human immunodeficiency virus [HIV], and avian influenza virus) and bacterial antigens and toxins (e.g., Staphylococcus aureus, Group A Streptococcus, and Escherichia coli). 13 In this study, we developed the immuno-PCR assay for PfHRP-2 and tested for the presence of CSF PfHRP-2 in CM patients. 14 We sought to test if there were elevated levels of CSF PfHRP2 compared with plasma in CM, and if elevated CSF PfHRP2 was associated with clinical outcome in this archival study of CM.…”

Section: Introductionmentioning

confidence: 99%

Quantification of Plasmodium falciparum Histidine-Rich Protein-2 in Cerebrospinal Spinal Fluid from Cerebral Malaria Patients

Mikita

Thakur

Anstey³

et al. 2014

The American Society of Tropical Medicine and Hygiene

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Abstract. A cerebrospinal fluid (CSF) biomarker for cerebral malaria (CM) has not been validated. We examined the detection, semiquantification, and clinical use of the Plasmodium falciparum histidine-rich protein-2 (PfHRP-2) as a parasite antigen biomarker for CM. The PfHRP-2 was detected in archival CSF samples from CM patients from Tanzania both by a newly developed sensitive and specific immuno-polymerase chain reaction (72 of 73) and by rapid diagnostic tests (62 of 73). The geometric mean PfHRP-2 CSF concentration was 8.76 ng/mL with no differences in those who survived (9.2 ng/mL), those who died (11

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“…47 Despite these features, ELISAs are not able to detect some antigens that are present at low concentrations. 48 ELISAs exist for Staphylococcus aureus detection in cases of bovine mastitis, 9 but the antibody titer does not correlate well with the amount of infecting bacteria. 62 Other…”

Section: Identification Of Pathogens Causing Intramammary Infectionmentioning

confidence: 99%

Technological advances in bovine mastitis diagnosis

2015

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Abstract. Bovine mastitis is an economic burden for dairy farmers and preventive control measures are crucial for the sustainability of any dairy business. The identification of etiological agents is necessary in controlling the disease, reducing risk of chronic infections and targeting antimicrobial therapy. The suitability of a detection method for routine diagnosis depends on several factors, including specificity, sensitivity, cost, time in producing results, and suitability for large-scale sampling of milk. This article focuses on current methodologies for identification of mastitis pathogens and for detection of inflammation, as well as the advantages and disadvantages of different methods. Emerging technologies, such as transcriptome and proteome analyses and nano-and microfabrication of portable devices, offer promising, sensitive methods for advanced detection of mastitis pathogens and biomarkers of inflammation. The demand for alternative, fast, and reliable diagnostic procedures is rising as farms become bigger. Several examples of technological and scientific advances are summarized which have given rise to more sensitive, reliable and faster diagnostic results.

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Immuno-PCR: a promising ultrasensitive diagnostic method to detect antigens and antibodies

Cited by 109 publications

References 55 publications

Enhanced Detection of Host Response Antibodies to Borrelia burgdorferi Using Immuno-PCR

Enhanced Detection of Host Response Antibodies to Borrelia burgdorferi Using Immuno-PCR

Quantification of Plasmodium falciparum Histidine-Rich Protein-2 in Cerebrospinal Spinal Fluid from Cerebral Malaria Patients

Technological advances in bovine mastitis diagnosis

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