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Start4All Pooled Testing Toolkit

  • The Start4All Pooled Testing Toolkit is an open access set of practical resources designed to support National TB Programmes and implementing partners in planning, introducing, evaluating, and scaling up pooled sputum testing for tuberculosis (TB). 

    The toolkit provides standard operating procedures, checklists, case studies, and instructional videos to guide consistent, safe, and effective implementation of pooled testing.

“In primary healthcare settings, pooled testing is a method that should be encouraged in low-income settings like ours… it’s economical and helps to screen more people with the same resources.” 

Cameroon KII1, Female, Programme Director

Toolkit Purpose

The purpose of the toolkit is to support National TB Programmes, especially in high TB burden settings in low- and middle-income countries, to adopt pooled sputum testing. It aims to: 

  • Expand access to molecular TB diagnosis by testing multiple sputum samples in a single cartridge or test. 
  • Reduce diagnostic costs without compromising test accuracy, supported by Start4All’s global evidence base.  
  • Support practical implementation, moving beyond proof of concept and enabling scale-up. 
  • Prepare for innovations, such as combining pooled testing with CAD guided triage and expanding to tongue swab specimens.
Start4All Pooled Sputum Testing Toolkit

Main Toolkit

This document contains –

  • Standard Operating Procedure (SOP)  – This SOP provides step‑by‑step guidance for implementing pooled sputum testing for TB, including how to prepare, combine, test, and interpret pooled samples using WHO‑recommended molecular platforms. It serves as a practical operational guide to ensure accurate, efficient, and safe use of pooled testing within routine laboratory workflows.
  • Frequently Asked Questions on pooled sputum testing for TB diagnosis – This document provides a clear set of frequently asked questions to support national TB programmes, clinicians, and laboratory teams in understanding and implementing pooled sputum testing for TB diagnosis. It outlines the purpose, operational steps, accuracy considerations, and common troubleshooting issues involved in introducing pooled testing into molecular TB workflows.
  • Site readiness checklist implementing pooled sputum testing – The checklist provides a structured way to assess whether a laboratory has the infrastructure, equipment, staff capacity, workflows, and quality assurance systems needed to begin pooled sputum testing using LC-aNAAT platforms. It enables programmes to determine if a site is fully ready, needs minor preparation, or is not yet prepared for implementation.
  • Two Case Studies: Case Study 1: When and why to use pooled testing – This case study provides practical guidance on when, why, and how to use pooled testing for TB diagnosis using LC-aNAAT technologies, drawing on extensive evidence and experience from the Start4All research programme. It outlines the conditions and settings where pooling is most effective, demonstrates its diagnostic performance and cost‑saving benefits, and offers implementation considerations for national TB programmes. The document also includes real-world lessons from large‑scale pooled‑testing rollout in Cameroon to support informed planning and scale‑up.
  • Case Study 2: How to implement and evaluate pooled testing – This case study provides practical, step-by-step guidance on how to implement routine pooled LC-aNAAT testing for TB, covering workflow design, biosafety, equipment needs, data linkage, and quality assurance requirements. It also outlines common implementation challenges, training needs, and lessons learned from the Start4All project to help programmes introduce pooled testing safely, efficiently, and with high diagnostic reliability.
Download main toolkit PDF

Poster

This poster provides a step by step visual guide for laboratories on how to perform pooled sputum testing for tuberculosis using LC-aNAAT.

It outlines specimen eligibility, preparation, pooling, result interpretation, and key quality and traceability requirements to support accurate and efficient implementation of pooling workflows.

Download poster (Powerpoint file)
Thumbnail image of a poster showing detailed instructions for Pooled Sputum Testing in TB diagnoses

Training presentation

This training presentation provides a practical, step‑by‑step guide for implementing pooled sputum testing for tuberculosis, covering workflow, eligibility, preparation, testing, interpretation, and quality assurance. It is designed to support laboratories and screening teams to apply pooled testing efficiently, maintain traceability, and ensure reliable diagnostic performance within the wider Pooling Toolkit.

Download training presentation (Powerpoint file)
Thumbnail image of a the first slide of a Powerpoint presentation with the title 'Pooled sputum testing'

Films

These short StopTB Partnership films introduce the concept of pooled sputum testing and provide clear, step-by-step guidance on how to perform the method. They are ideal for training staff and supporting consistent implementation across teams.

What is pooled testing and why is it relevent?

How to perform pooled testing for TB

In English:

In Spanish:

In French: