Lateral Flow Immunoassay for Naked Eye Detection of Mycobacterium tuberculosis

Abstract

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis. Detection and control of infectious diseases is a major problem, especially in developing countries. Lateral flow immunoassay (LFIA) has been introduced as a handheld immunoassay-based point-of-care platform for an automated detection of TB. The CFP10-ESAT6 antigen of M. tuberculosis was used as the target in early detection of TB using LFIA strip-based POC strategy. An interesting platform based on optical signals is implemented as a colour change in the detection area that is visible to the naked eye. The gold nanoparticles (AuNPs) were used as the colour probe for the detection of a target of interest. The high-resolution transmission electron microscopy (HRTEM) image and ultraviolet-visible spectrophotometer (UV-Vis) analysis confirmed that the synthesized AuNPs were appropriate for the immunoassay designed. The platform consists of AuNPs conjugated with specific antibodies (Ab) to capture the antigen of M. tuberculosis. Under the capillary effect, sandwich immunoreactions of AuNP-Ab-antigen were performed on the test pad of the immunostrip, which can be observed by the colour signal on the test line of the strip with a short assay time. Furthermore, the newly developed biosensor was utilized in CFP10-ESAT6 antigen detection in human sputum specimens with satisfactory results. The characteristic coloured bands enable visual detection (naked eye) of target analyte without instrumentation. This noninvasive diagnose system which is sputum-based detection could provide user-friendly and affordable diagnostic tests in developing countries.

Detection of HIV and Mycobacterium tuberculosis among University Students in Port Harcourt, Nigeria

“…Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Nazifah et al, 2020), and Africa is facing the worst TB epidemic since the advent of the antibiotic era. Driven by a generalized human immunodeficiency virus (HIV) epidemic and compounded by weak health care systems, inadequate laboratories, and conditions that promote transmission of infection, this devastating situation has steadily worsened, exacerbated by the emergence of drug-resistant strains of Mycobacterium tuberculosis (Chaisson and Martinson, 2008; Okonko et al, 2012a,b,c).…”

“…Driven by a generalized human immunodeficiency virus (HIV) epidemic and compounded by weak health care systems, inadequate laboratories, and conditions that promote transmission of infection, this devastating situation has steadily worsened, exacerbated by the emergence of drug-resistant strains of Mycobacterium tuberculosis (Chaisson and Martinson, 2008; Okonko et al, 2012a,b,c). Detection and control of infectious diseases is a major problem, especially in developing countries (Nazifah et al, 2020).…”

Detection of Bacterial and Viral Pathogens Using Photonic Point-of-Care Devices

“…LFIAs generally consists of a sample loading pad, absorbent pad, conjugate pad, a test line, and a control line on a membrane (commonly used nitrocellulose membrane). One example of the LFIA is the recently developed tuberculosis detection, where the sample is deposited on the loading pad and flows laterally to meet the conjugate pad that contains immobilized gold nanoparticles (AuNPs) tagged with antibody (Ab) that specifically captures the CFP10-ESAT6 antigen of M. tuberculosis in the sample [ 46 ]. The AuNP-Ab-Antigen complex then flows along the membrane laterally due to the self-capillary action of the membrane and meets the test line, which has a second antibody that captures the AuNP-Ab-Antigen complex, resulting in a colored line.…”

Preparasi Strip Imunokromatografi Koloid Emas untuk Deteksi Cepat Aeromonas hydrophila

“…Keterangan: A. Nilai absorbansi koloid emas dengan Koloid emas hasil sintesis dengan metode ini digunakan sebagai label pada strip imunokromatografi yang dikembangkan. Karakteristik koloid emas dengan puncak serapan pada panjang gelombang 525 nm dengan ukuran partikel 30 nm telah dilaporkan (Rahme et al, 2013;Zeng et al, 2014;Berlina et al, 2019) dan banyak peneliti menggunakan koloid emas dengan ukuran partikel tersebut sebagai label pada pembuatan strip imunokromatografi (Sun et al, 2012;Liu et al, 2014;Ariffin et al, 2020). Koloid emas dengan ukuran partikel 31,88 nm pada penelitian ini memberikan kestabilan yang baik ketika dikonjugasikan dengan jumlah optimum antibodi anti A. hydrophila.…”

“…Jumlah antibodi dibawah 12,5 µg terjadi perubahan warna dari merah ke biru, yang mengindikasikan bahwa koloid emas tidak dapat distabilkan pada jumlah tersebut. Beberapa penelitian sebelumnya (Zhou et al, 2014;Wen-de et al, 2017;Ariffin et al, 2020) antibodi yang diaplikasikan pada pembuatan konjugat adalah 1,2 kali jumlah minimum antibodi dalam mestabilkan koloid emas. Di dalam penelitian ini, konjugat yang diaplikasikan dengan 1,2 kali jumlah minimum antibodi memberikan warna yang tipis dan hampir tidak terlihat dengan mata telanjang pada garis uji (data tidak ditampilkan).…”