-1::1
Simple Hit Counter
Skip to content

Products

Solutions

×
×
Sign In

ES

EN - EnglishCN - 简体中文DE - DeutschES - EspañolKR - 한국어IT - ItalianoFR - FrançaisPT - Português do BrasilPL - PolskiHE - עִבְרִיתRU - РусскийJA - 日本語TR - TürkçeAR - العربية
Sign In Start Free Trial

RESEARCH

JoVE Journal

Peer reviewed scientific video journal

Behavior
Biochemistry
Bioengineering
Biology
Cancer Research
Chemistry
Developmental Biology
View All
JoVE Encyclopedia of Experiments

Video encyclopedia of advanced research methods

Biological Techniques
Biology
Cancer Research
Immunology
Neuroscience
Microbiology
JoVE Visualize

Visualizing science through experiment videos

EDUCATION

JoVE Core

Video textbooks for undergraduate courses

Analytical Chemistry
Anatomy and Physiology
Biology
Calculus
Cell Biology
Chemistry
Civil Engineering
Electrical Engineering
View All
JoVE Science Education

Visual demonstrations of key scientific experiments

Advanced Biology
Basic Biology
Chemistry
View All
JoVE Lab Manual

Videos of experiments for undergraduate lab courses

Biology
Chemistry

BUSINESS

JoVE Business

Video textbooks for business education

Accounting
Finance
Macroeconomics
Marketing
Microeconomics

OTHERS

JoVE Quiz

Interactive video based quizzes for formative assessments

Authors

Teaching Faculty

Librarians

K12 Schools

Biopharma

Products

RESEARCH

JoVE Journal

Peer reviewed scientific video journal

JoVE Encyclopedia of Experiments

Video encyclopedia of advanced research methods

JoVE Visualize

Visualizing science through experiment videos

EDUCATION

JoVE Core

Video textbooks for undergraduates

JoVE Science Education

Visual demonstrations of key scientific experiments

JoVE Lab Manual

Videos of experiments for undergraduate lab courses

BUSINESS

JoVE Business

Video textbooks for business education

OTHERS

JoVE Quiz

Interactive video based quizzes for formative assessments

Solutions

Authors
Teaching Faculty
Librarians
K12 Schools
Biopharma

Language

Spanish

EN

English

CN

简体中文

DE

Deutsch

ES

Español

KR

한국어

IT

Italiano

FR

Français

PT

Português do Brasil

PL

Polski

HE

עִבְרִית

RU

Русский

JA

日本語

TR

Türkçe

AR

العربية

    Menu

    JoVE Journal

    Behavior

    Biochemistry

    Bioengineering

    Biology

    Cancer Research

    Chemistry

    Developmental Biology

    Engineering

    Environment

    Genetics

    Immunology and Infection

    Medicine

    Neuroscience

    Menu

    JoVE Encyclopedia of Experiments

    Biological Techniques

    Biology

    Cancer Research

    Immunology

    Neuroscience

    Microbiology

    Menu

    JoVE Core

    Analytical Chemistry

    Anatomy and Physiology

    Biology

    Calculus

    Cell Biology

    Chemistry

    Civil Engineering

    Electrical Engineering

    Introduction to Psychology

    Mechanical Engineering

    Medical-Surgical Nursing

    View All

    Menu

    JoVE Science Education

    Advanced Biology

    Basic Biology

    Chemistry

    Clinical Skills

    Engineering

    Environmental Sciences

    Physics

    Psychology

    View All

    Menu

    JoVE Lab Manual

    Biology

    Chemistry

    Menu

    JoVE Business

    Accounting

    Finance

    Macroeconomics

    Marketing

    Microeconomics

Start Free Trial
Loading...
Home
JoVE Journal
Biochemistry
Secuenciación de amplicones utilizando las tecnologías de secuenciación de lectura larga
Secuenciación de amplicones utilizando las tecnologías de secuenciación de lectura larga
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Amplicon Sequencing using the Long-Read Sequencing Technologies

Secuenciación de amplicones utilizando las tecnologías de secuenciación de lectura larga

Full Text
606 Views
08:57 min
August 29, 2025

DOI: 10.3791/68370-v

Morwasehla Modjadji1, Brendon Coenrad Mann1, Johannes Loubser1, Jennifer Williams1, Janré Steyn1, Elizabeth Maria Streicher1, Melanie Grobbelaar1, Robin Mark Warren1

1Department of Science and Innovation - National Research Foundation Centre of Excellence for Biomedical Tuberculosis Research, South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences,Stellenbosch University

AI Banner

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study investigates the use of portable long-read sequencing for determining drug-resistant tuberculosis (TB) susceptibility in resource-limited settings. The protocol aims to provide accurate results comparable to gold standard methods, enhancing accessible TB diagnostics.

Key Study Components

Area of Science

  • Microbiology
  • Genomics
  • Infectious Diseases

Background

  • Drug-resistant tuberculosis is a significant global health challenge.
  • Traditional diagnostic methods may be limited in resource-poor settings.
  • Long-read sequencing offers potential advantages in accuracy and turnaround time.
  • Bioinformatics expertise is often a barrier to effective diagnostics.

Purpose of Study

  • To evaluate the effectiveness of long-read sequencing for TB diagnostics.
  • To provide a comprehensive resistance profile from clinical samples.
  • To simplify the workflow for drug-resistance detection.

Methods Used

  • Targeted next-generation sequencing of 18 drug-resistance regions.
  • Utilization of a tuberculosis-specific bioinformatics pipeline.
  • Sample preparation involving precise measurement of amplicons.
  • Adjustment of sample volumes for optimal sequencing results.

Main Results

  • Long-read sequencing demonstrated accuracy comparable to gold standards.
  • Quicker turnaround times were achieved compared to traditional methods.
  • The protocol facilitated comprehensive TB diagnostics.
  • Enhanced outbreak tracking was possible in resource-limited environments.

Conclusions

  • Portable long-read sequencing can improve TB diagnostics in low-resource settings.
  • The method reduces the need for extensive bioinformatics expertise.
  • This approach has the potential to transform TB management and control strategies.

Frequently Asked Questions

What is the significance of drug-resistant tuberculosis?
Drug-resistant tuberculosis poses a major public health threat, complicating treatment and control efforts.
How does long-read sequencing compare to traditional methods?
Long-read sequencing offers higher accuracy and faster results, making it suitable for resource-limited settings.
What are the challenges in TB diagnostics?
Challenges include limited access to advanced technologies and the need for specialized bioinformatics skills.
Can this method be used in clinical settings?
Yes, the protocol is designed to be implemented directly from clinical samples.
What are the implications for public health?
Improved diagnostics can lead to better management of TB outbreaks and more effective treatment strategies.

Este protocolo se optimizó para la secuenciación profunda dirigida de 18 regiones de resistencia a los medicamentos en Mycobacterium tuberculosis utilizando una plataforma de secuenciación de lectura larga, seguida de un análisis con una línea de bioinformática específica para la tuberculosis diseñada para datos de lectura larga.

Investigo si la secuenciación portátil de lectura larga puede proporcionar resultados de susceptibilidad resistente a la tuberculosis en entornos con recursos limitados, con una precisión comparable al método de estándares de oro, avanzando en diagnósticos de tuberculosis accesibles y de alta calidad. Utilizar secuenciación dirigida de nueva generación como herramienta diagnóstica para la tuberculosis resistente a medicamentos, ofreciendo un perfil completo de resistencia directamente de la muestra clínica sin conocimientos en bioinformática. Nuestro protocolo, que utiliza secuenciación de lecturas largas, tiene el potencial de proporcionar tiempos de respuesta más rápidos, un flujo de trabajo más sencillo que en la detección de resistencia en tiempo real, facilitando diagnósticos completos de TB y mejorando el seguimiento de brotes en entornos con recursos limitados y una infraestructura mínima.

Para empezar, calcula 100 nanogramos de cada muestra de amplicón en función de la concentración y transfiere el volumen requerido a un tubo PCR limpio. Para determinar la masa total de amplicones en la muestra, se utiliza la fórmula dada. Ajusta el volumen total de cada muestra a 12,5 microlitros usando agua libre de nucleasa.

View the full transcript and gain access to thousands of scientific videos

View the full transcript and gain access to thousands of scientific videos

Sign In Start Free Trial

Explore More Videos

Bioquímica Número 222

Related Videos

Solo lectura y pareadas final Sec ARNm bibliotecas de Illumina de 10 nanogramos ARN total

14:49

Solo lectura y pareadas final Sec ARNm bibliotecas de Illumina de 10 nanogramos ARN total

Related Videos

39.7K Views

La secuenciación de próxima generación de 16S ARN ribosomal genes Amplicones

10:24

La secuenciación de próxima generación de 16S ARN ribosomal genes Amplicones

Related Videos

84.8K Views

Extremo 3' secuencia biblioteca elaboración con Sec2 A

12:01

Extremo 3' secuencia biblioteca elaboración con Sec2 A

Related Videos

11.1K Views

Muy largo leer la secuencia para el análisis de la DNA de Genomic entero

10:34

Muy largo leer la secuencia para el análisis de la DNA de Genomic entero

Related Videos

24.1K Views

Secuenciación de ARNm de sangre entera utilizando secuenciación de nanoporos

11:26

Secuenciación de ARNm de sangre entera utilizando secuenciación de nanoporos

Related Videos

14.9K Views

Ensamblaje híbrido del genoma de novo para la generación de genomas completos de bacterias urinarias utilizando tecnologías de secuenciación de lectura corta y larga

12:08

Ensamblaje híbrido del genoma de novo para la generación de genomas completos de bacterias urinarias utilizando tecnologías de secuenciación de lectura corta y larga

Related Videos

5.9K Views

Cribado de fármacos basado en transcriptómica rentable

06:40

Cribado de fármacos basado en transcriptómica rentable

Related Videos

1.8K Views

Perfil espacial completo de transcriptomas independientes de la especie a través de Stereo-seq

10:22

Perfil espacial completo de transcriptomas independientes de la especie a través de Stereo-seq

Related Videos

721 Views

Rab10 detección de fosforilación por la actividad de LRRK2 mediante SDS-PAGE con una etiqueta de enlace de fosfato

08:55

Rab10 detección de fosforilación por la actividad de LRRK2 mediante SDS-PAGE con una etiqueta de enlace de fosfato

Related Videos

16.2K Views

Definición acompañante Regulación Redox actividad de Hsp33 y mapeo de cambios conformacionales en la Hsp33 mediante espectrometría de masas de intercambio hidrógeno-deuterio

10:24

Definición acompañante Regulación Redox actividad de Hsp33 y mapeo de cambios conformacionales en la Hsp33 mediante espectrometría de masas de intercambio hidrógeno-deuterio

Related Videos

9.3K Views

JoVE logo
Contact Us Recommend to Library
Research
  • JoVE Journal
  • JoVE Encyclopedia of Experiments
  • JoVE Visualize
Business
  • JoVE Business
Education
  • JoVE Core
  • JoVE Science Education
  • JoVE Lab Manual
  • JoVE Quizzes
Solutions
  • Authors
  • Teaching Faculty
  • Librarians
  • K12 Schools
  • Biopharma
About JoVE
  • Overview
  • Leadership
Others
  • JoVE Newsletters
  • JoVE Help Center
  • Blogs
  • JoVE Newsroom
  • Site Maps
Contact Us Recommend to Library
JoVE logo

Copyright © 2026 MyJoVE Corporation. All rights reserved

Privacy Terms of Use Policies
WeChat QR code