Banca de DEFESA: ANA CLAUDIA NEGRET SCALIA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : ANA CLAUDIA NEGRET SCALIA
DATE: 25/11/2022
TIME: 14:30
LOCAL: Plataforma Teams
TITLE:

ACURÁCIA DA REAÇÃO EM CADEIA DE POLIMERASE EM TEMPO REAL PARA O DIAGNÓSTICO DA HANSENÍASE EM DIFERENTES FASES DO SANGUE PERIFÉRICO: UM MODELO DIAGNÓSTICO E DE DISSEMINAÇÃO DO Mycobacterium leprae

KEY WORDS:

Mycobacterium leprae, qPCR, leprosy, sensibility, quantification, public health

PAGES: 71
BIG AREA: Ciências da Saúde
AREA: Medicina
SUMMARY:

It is believed that leprosy is the oldest infectious and contagious disease of the humanity and until today it is a public health problem worldwide. According to WHO, 161 countries reported data on the disease in 2019 and the total of new cases reached 202.256. Brazil, a country included in the priority group for the disease control, in the same year reported 27.864 incidents, of which 2.351 were classified as Disability Degree 2 and 1.545 were child cases. Since the diagnosis of the disease is exclusively clinical and the laboratory tests that exist today are used only as an aid tool, it is important to develop a less invasive tool, a less costely for the public coffers and that allows the confirmation of the clinical condition reported by the doctor. This study proposes the detection and quantification of Mycobacterium leprae in the different components of peripheral whole blood through real-time Polymerase Chain Reaction (qPCR). The study population consisted of leprosy suspected patients, treated at the University Hospital of Brasília, during the one-year period between June 2000 and June 2021, totaling 111 patients, of which 55 were positively diagnosed and 56 were controls. After obtaining the serum, plasma, red blood cells and peripheral blood mononuclear cells, they were subjected to DNA extraction using a commercial Purelink Genomic kit from Thermofisher. For the standarization of the positive controls and the reaction of the quantification standard curve, the RLEP genetic target of the mycobacterial genome was made. The qPCR was performed qualitatively and quantitatively using the primers RLEPBR F -5’ – CTTGCACCATTTCTGCCGCT – 3’ e RLEPBR R – 5’ – TGCGCTAGAAGGTTGCCGTA – 3’. Of the participants clinically diagnosed with leprosy, 52,73% were female, the average age was 44.33 years and none were household contacts. Of the control participants, 50% were male, the average age was 45.11 and 48 were household contacts. By the classification of the clinical condition, 1 participant was indeterminate, 5 were tuberculoid, 8 borderline-tuberculoid, 18 borderline, 11 borderlinelepromatous and 12 lepromatous. As for the disability degree, 17 were classified as degree 1, 15 patients as degree 2 and 23 with none degree. As for the biomolecular results, the sensitivity was low corresponding to 11.54% in serum, 7.31% in plasma, 9.26% in peripheral blood mononuclear cells, 9.80% in red blood cells and 38.18% in smear microscopy. Among paucibacillary patients, the highest sensitivity was found in both serum and red blood cells and among multibacillary patients the highest sensitivity was found in serum. The low sensitivity found in the results suggests that peripheral blood is not a consistent material for diagnosing leprosy.

BANKING MEMBERS:
Externo à Instituição - THAIS MONTEZUMA
Externo à Instituição - ALEXANDRE CASIMIRO DE MACEDO - MS
Presidente - 2676451 - CIRO MARTINS GOMES
Externo à Instituição - DANIEL HOLANDA BARROSO - HUB