Evaluation and comparison of automated and manual ELISA for diagnosis of chronic pulmonary aspergillosis (CPA) in Indonesia
DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE
Authors: Setianingrum, Findra; Rozaliyani, Anna; Syam, Ridhawati; Adawiyah, Robiatul; Tugiran, Mulyati; Sari, Cut Yulia, I; Burhan, Erlina; Wahyuningsih, Retno; Rautemaa-Richardson, Riina; Denning, David W.
Abstract
Pulmonary tuberculosis (TB) is one of the common risk factors for chronic pulmonary aspergillosis (CPA). A positive Aspergillus IgG is a key element of the diagnosis of CPA but this has not been studied in Indonesia. We conducted studies with patients at the end of TB therapy in Indonesia. We performed receiver operating curve (ROC) analysis to determine the optimum cutoff of the Aspergillus-specific IgG level (Immulite and Dynamiker ELISA) in those patients who met criteria of CPA in relation to control groups. In 203 TB patients, 26 (13%) patients had clinical and radiological features of CPA. We derived optimum cutoffs for Immulite Aspergillus-specific IgG of 11.5 mg/L and Dynamiker anti-galactomannan IgG of 106.8 AU/mL (sensitivity 89% and 83%, specificity 78% and 51%, respectively). The currently accepted Aspergillus-specific IgG cutoff of Immulite and Dynamiker assays for CPA diagnosis may require slight adjustment for the Indonesian population. (C) 2020 Elsevier Inc. All rights reserved.
Immunoprotection against lethal effects of Crotalus durissus snake venom elicited by synthetic epitopes trapped in liposomes
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Authors: Vaz de Melo, Patricia D.; Lima, Sabrina de Almeida; Araujo, Priscila; Santos, Raissa Medina; Gonzalez, Edgar; Belo, Andreza Alves; Machado-de-Avila, Ricardo A.; Costal-Oliveira, Fernanda; Soccol, Vanete T.; Guerra-Duarte, Clara; Rezende, Leonides; Chavez-Olortegui, Carlos
Abstract
Snakebites caused by Crotalus genus are the second most frequent in Brazil. Crotoxin is a beta-neurotoxin responsible for the main envenomation effects of Crotalus biting, while crotamine immobilizes the animal hind limbs, contributing to prey immobilization and to envenoming symptoms. As crotoxin and crotamine represent about 90% of Crotalus venom dry weight, these toxins are of great importance for antivenom therapy. In this sense, knowledge regarding the antigenicity/immunogenicity at the molecular level of these toxins can provide valuable information for the improvement of specific antivenoms. Therefore, the aims of this study are the identification of the B-cell epitopes from crotoxin and crotamine; and the characterization of the neutralizing potency of antibodies directed against the corresponding synthetic epitopes defined in the current study. Linear B-cell epitopes were identified using the Spot Synthesis technique probed with specific anti -C. d. terrificus venom horse IgG. One epitope of crotamine (F(12)PKEKICLPPSSDFGKMDCRW(32)) and three of crotoxin (L(10)LVGVEGHLLQFNKMIKFETR(30); Y(43)CGWGGRGRPKDATDRCCFVH(63) and T(118)YKYGYMFYPDSRCRGPSETC(138)) were identified. After synthesis in their soluble form, the peptides mixture correspondent to the mapped epitopes was entrapped in liposomes and used as immunogens for antibody production in rabbits. Anti-synthetic peptide antibodies were able to protect mice from the lethal activity of C. d. terrificus venom. (C) 2020 Elsevier B.V. All rights reserved.