Evaluation of the Western blotting method for the diagnosis of congenital toxoplasmosis

Capobiango, Jaqueline Dario; Monica, Thaís Cabral; Ferreira, Fernanda Pinto; Mitsuka‐Breganó, Regina; Navarro, Italmar Teodorico; Garcia, João Luís; Reiche, Edna Maria Vissoci

doi:10.1016/j.jped.2016.02.014

Cited by 22 publications

(3 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Among the selected studies, the use of samples such as serum, whole blood, colostrum and amniotic fluid was observed, and these were associated with different methods of detection of T. gondii and its respective markers. Table 1 presents the general characteristics of the selected articles [6][7][8][9][10][11][12][13][14][15][16][17][18] .…”

Section: General Characteristics Of the Study And Total Resultsmentioning

confidence: 99%

Molecular and serological diagnosis of toxoplasmosis: a systematic review and meta-analysis

Souza

Siqueira

Ribeiro

et al. 2023

Rev. Inst. Med. trop. S. Paulo

View full text Add to dashboard Cite

Toxoplasmosis is an infection of vast worldwide distribution whose etiologic agent is Toxoplasma gondii. This disease can cause problems ranging from mild symptoms to serious conditions, such as encephalitis, miscarriage and blindness. Therefore, it is of utmost importance to perform a diagnosis with reproducible techniques in order to obtain a good prognosis. The aim of this review was to analyze the efficiency of toxoplasmosis diagnostic techniques based on sensitivity and specificity results. Five research platforms in English language were used (Eric, Elsevier, Google Scholar, PubMed and SciELO), which contained data on the diagnosis of toxoplasmosis. The search and selection were performed for studies published prior to June 2021. The search resulted in the inclusion of 13 articles published from 2005 to 2020. The data revealed the use of different samples in the standardization of techniques such as serum, total blood, colostrum and amniotic fluid. The flow cytometry, lateral flow immunoassay and qPCR techniques showed 100% sensitivity, whereas the ELISA, western blotting, qPCR and RE-LAMP techniques achieved 100% specificity. Significantly, the qPCR and LAMP techniques were more accurate when the likelihood ratio was assessed. The meta-analysis identified that ISAGA and western blotting have low sensitivity values and LIASON, ELFA and ELISA, using a silica bioconjugate, also have low specificity values. It was noted that a wide range of methods have high values of sensitivity and specificity. Therefore, the choice of the method will be based on the conditions and its financial viability.

show abstract

Section: General Characteristics Of the Study And Total Resultsmentioning

confidence: 99%

Molecular and serological diagnosis of toxoplasmosis: a systematic review and meta-analysis

Souza

Siqueira

Ribeiro

et al. 2023

Rev. Inst. Med. trop. S. Paulo

View full text Add to dashboard Cite

show abstract

“…An implemented protocol for an early CT diagnosis may also be achieved with more specific blood tests. Capobiango JD et al [ 7 ] have described, an immunoblot test for the comparison of mother and infant IgG and IgM immunological profiles at birth or shortly after. Mahmoudi S et al characterized an interferon-gamma release assay (IGRA) where an early CT diagnosis was achieved by stimulating T-cells from infected neonates with T. gondii antigens [ 8 ] .…”

Section: Discussionmentioning

confidence: 99%

Specific antibody titer decay in neonates prenatally exposed to Toxoplasma gondii and their mothers

et al. 2022

View full text Add to dashboard Cite

Background For infants exposed in utero to Toxoplasma gondii, current guidelines recommend monitoring the specific antibody titer until 12 months of age. In this study, we investigated the antibody titer decay in the mother-infant dyad. Methods This is a single center, population-based cohort study of neonates referred for prenatal exposure to Toxoplasma gondii from January 2014 to December 2020. All infants underwent clinical, laboratory, and instrumental investigation for at least 12 months. Results A total of 670 eligible neonates were referred to the Perinatal Infection Unit of the University Federico II of Naples. 636 (95%) completed the serological follow up until 12 months. Specific IgG antibodies negativization occurred in 628 (98.7%) within 5 months. At 9 and 12 months, all patients had negative IgG. An initial neonatal IgG antibody titer ≥ 200 IU/ml was associated with a longer time to negativization (184 [177.5;256] days when above threshold vs. 139.5 [101;179] days when below it; p < 0.001). Maternal IgG antibody titer ≥ 200 IU/ml at childbirth was also associated to delayed time to negativization in the infant (179 [163;184] days above the threshold vs 125 [96.8;178] days below it; p < 0.001). Specific antibody negativization was irreversible in all patients. Conclusions Lower anti-Toxoplasma antibody titers detected at birth in the mother-infant-dyad lead to an earlier and irreversible negativization. This information allows for customisation of the infant follow up program and avoids invasive and expensive tests.

show abstract

“…To overcome diagnostic issues of lower sensitivity and specificity, the immunoreactivity to Mycobacterium tuberculosis antigens was performed by western blotting [78]. Furthermore, western blotting was performed for the early and sensitive diagnosis of congenital toxoplasmosis [79] and was employed for rapid and sensitive serological diagnosis of a serious infectious disease paracoccidioidomycosis (PCM) [80]. Using immunoblotting, a new subgroup of human lymphotropic retroviruses (HTLV), was detected in patients with the acquired immunodeficiency syndrome (AIDS) [81].…”

Section: Relationship Between Improved Western Blot Methods and Its Cmentioning

confidence: 99%

Protein purification and analysis: next generation Western blotting techniques

Mishra

Tiwari

Gomes

2017

Expert Review of Proteomics

168

View full text Add to dashboard Cite

Western blotting is one of the most commonly used techniques in molecular biology and proteomics. Since western blotting is a multistep protocol, variations and errors can occur at any step reducing the reliability and reproducibility of this technique. Recent reports suggest that a few key steps, such as the sample preparation method, the amount and source of primary antibody used, as well as the normalization method utilized, are critical for reproducible western blot results. Areas covered: In this review, improvements in different areas of western blotting, including protein transfer and antibody validation, are summarized. The review discusses the most advanced western blotting techniques available and highlights the relationship between next generation western blotting techniques and its clinical relevance. Expert commentary: Over the last decade significant improvements have been made in creating more sensitive, automated, and advanced techniques by optimizing various aspects of the western blot protocol. New methods such as single cell-resolution western blot, capillary electrophoresis, DigiWest, automated microfluid western blotting and microchip electrophoresis have all been developed to reduce potential problems associated with the western blotting technique. Innovative developments in instrumentation and increased sensitivity for western blots offer novel possibilities for increasing the clinical implications of western blot.

show abstract

Evaluation of the Western blotting method for the diagnosis of congenital toxoplasmosis

Abstract: The IgG-WB showed greater sensitivity than the detection of IgM anti-T. gondii; therefore, it can be used for the diagnosis of congenital toxoplasmosis in association with other congenital infection markers.

Cited by 22 publications

References 20 publications

Molecular and serological diagnosis of toxoplasmosis: a systematic review and meta-analysis

Molecular and serological diagnosis of toxoplasmosis: a systematic review and meta-analysis

Specific antibody titer decay in neonates prenatally exposed to Toxoplasma gondii and their mothers

Protein purification and analysis: next generation Western blotting techniques

Contact Info

Product

Resources

About