WHO is developing its first global guidelines on multiplex testing for HIV, hepatitis, and STIs, offering lab leaders a framework to optimize diagnostics, streamline workflows, and lead the shift toward integrated, multi-disease testing.
As diagnostic technologies rapidly evolve and healthcare systems shift toward integrated service delivery, laboratory leaders are at the forefront of implementing efficient, multi-disease testing strategies.
In a landmark move, the World Health Organization (WHO) has convened a Guideline Development Group (GDG) to develop its first-ever evidence-based recommendations on multiplex testing, a method that enables simultaneous detection of HIV, viral hepatitis, and STIs from a single sample. This initiative will establish foundational principles for integration, resource prioritization, and diagnostic efficiency, offering lab professionals critical guidance to shape the future of testing across diseases.
The GDG, comprised of international experts and stakeholders, will provide evidence-based recommendations to support integrated, people-centered diagnostic strategies.
As global health systems shift toward more integrated service delivery, multiplex testing is emerging as a practical tool for increasing diagnostic access, streamlining care, and maximizing limited resources, particularly in low- and middle-income countries, where testing gaps remain significant.
“Access to timely and accurate diagnostic testing is essential for the prevention, detection, and management of HIV, viral hepatitis and STIs,” WHO stated in its announcement. “Multiplex testing has emerged as a promising strategy to improve efficiency, expand testing coverage across diseases, and enhance cost-effectiveness.”
A First-of-Its-Kind Guideline
This is the first WHO guideline to explicitly address multiplex testing using the most up-to-date evidence. While the focus will be on HIV, viral hepatitis, and STIs, the guidelines aim to establish critical principles for integration that can be extended to other disease areas over time.
“This guideline will provide critical principles for integration that drive public health impact and chart the course for further multi-disease testing approaches,” WHO noted.
Photo credit: “World Health Organization Flag” by United States Mission Geneva is licensed under CC BY 2.0. To view a copy of this license, visit https://creativecommons.org/licenses/by/2.0/?ref=openverse.
The GDG will address both provider-based testing and self-testing, recognizing the growing role of community-led and patient-driven diagnostics. Key issues on the agenda include how to prioritize limited testing resources, ensure quality assurance, and optimize public health outcomes through integrated diagnostic models.
Guideline Development Group: Global, Inclusive, Independent
The GDG is composed of members from all WHO regions, selected based on their technical expertise, field experience, and perspectives as either implementers or members of affected communities. Importantly, all members participate in their individual capacities and not as representatives of any affiliated organizations.
“In accordance with WHO guidelines for developing recommendations, the GDG is composed of members from all WHO regions, serving in their individual capacities,” the organization explained. “Members do not receive financial compensation for their contributions to this process.”
The GDG includes program managers, healthcare providers, researchers, and community advocates, reflecting the full spectrum of stakeholders involved in diagnostic service delivery. Their diverse backgrounds are intended to ensure that the guidelines are evidence-informed, practical, and contextually relevant across settings.
A virtual meeting is scheduled for November 4-5, 2025, during which the group will discuss evidence, identify priority recommendations, and finalize key principles around multiplex testing and integration.
WHO Opens Public Comment Period
To promote transparency and inclusivity, WHO has opened a public comment period and is inviting feedback on the composition of the GDG. Stakeholders, organizations, and individuals can review member biographies and submit comments via email to hiv-aids@who.int by September 29, 2025.
The upcoming guideline represents a significant step in advancing integrated diagnostics and expanding access to care, particularly in resource-limited settings. By combining clinical evidence with real-world insights, WHO aims to provide countries and implementers with practical, scalable recommendations that improve testing coverage and disease detection at all levels of the health system.
As testing technologies evolve and the demand for multi-disease platforms grows, this guideline may serve as a blueprint for future diagnostic integration efforts, laying the foundation for efficient, patient-centered, and cost-effective care delivery worldwide.
A UCLA microbiology lab used whole-genome sequencing to trace a carbapenem-resistant Pseudomonas outbreak to a single ICU sink, revealing how biofilm and plumbing can silently harbor superbugs.
A routine culture from an ICU patient at UCLA Health sparked an investigation that ultimately uncovered a silent, domestic outbreak of a highly resistant strain of Pseudomonas aeruginosa. The discovery was led by the Molecular Microbiology and Pathogen Genomics Laboratory and highlights the critical role clinical laboratories play in outbreak detection, antimicrobial resistance surveillance, and environmental tracking.
The initial isolate appeared typical: P. aeruginosa, a common hospital-associated pathogen. But further analysis revealed something more troubling, the presence of NDM-1 (New Delhi metallo-β-lactamase), an enzyme that breaks down carbapenems and other powerful beta-lactam antibiotics, rendering them ineffective.
“This was the first time we’d ever seen an NDM-1-producing Pseudomonas strain in our hospital—and in a patient with no international travel,” said Shangxin Yang, PhD, director of UCLA Health’s Molecular Microbiology and Pathogen Genomics Laboratory.
Shangxin Yang, PhD, director of UCLA Health’s Molecular Microbiology and Pathogen Genomics Laboratory noted, “While NDM-1 is prevalent in Asia, Europe and the Middle East, it remains rare in the United States. That’s when we knew this wasn’t imported. This was something domestic—and very concerning.” (Photo credit: UCLA)
Sporadic Cases, Elusive Source
Over the next 18 months, seven additional patients were identified with the same rare resistance pattern. The cases were sporadic—spread across time and units—and did not follow conventional outbreak patterns, complicating source identification.
In collaboration with UCLA Health’s infection prevention team, the lab launched a detailed investigation. Routine epidemiologic methods failed to identify commonalities between the cases. Shared equipment, staffing patterns, and care protocols were ruled out. With limited leads, the microbiology team turned to whole-genome sequencing (WGS).
Whole-Genome Sequencing Connects the Dots
WGS became the turning point. By sequencing all eight patient isolates and comparing them to environmental samples, Yang’s lab determined that seven of the eight clinical isolates and two environmental strains shared an almost identical genomic profile. Only one isolate, from a patient previously treated in Iran, was genetically distinct.
“Whole-genome sequencing gave us the clarity we needed,” said Yang. “It allowed us to move from hypothesis to high-resolution confirmation—pinpointing the genetic relatedness of these organisms with certainty.”
The team had uncovered a clonal outbreak of NDM-1-producing P. aeruginosa, likely stemming from a single environmental reservoir.
Unexpected Reservoir: An ICU Sink
During a third round of environmental testing, the lab isolated the same NDM-1-producing strain from a contaminated sink drain and P-trap in one ICU room. Notably, two of the eight patients had been admitted to that room more than a year apart.
The persistence of the organism was attributed to biofilm formation in the sink plumbing. Pseudomonas is known for forming robust biofilms that adhere to moist surfaces and resist standard disinfection methods.
“This wasn’t just about surface contamination,” said Yang. “This was a deeply embedded reservoir that conventional cleaning protocols couldn’t touch.”
Lab-Driven Response and Mitigation
Once the lab identified the environmental source, targeted interventions were put in place:
Weekly disinfection of ICU sinks using Virasept, a biofilm-effective agent
Plumbing replacement, including P-trap components known to harbor persistent biofilms
Engineering modifications to faucet angles to reduce splash-back and droplet spread
Expanded environmental surveillance to monitor other sinks for colonization
The lab continued to monitor the situation post-intervention, and no further cases of NDM-1-producing P. aeruginosa have been identified since the changes were implemented.
Lessons Learned
This case reinforces the value of whole-genome sequencing in resolving complex outbreaks, linking patient isolates to an environmental source that traditional methods missed. It highlights the need to include plumbing and other biofilm-prone areas in environmental sampling. Most importantly, it shows how microbiology labs through genomic, phenotypic, and molecular tools can lead outbreak investigations, especially when paired with strong cross-department collaboration.
“This is a clear example of the power of the clinical lab when genomic tools and environmental surveillance are used strategically,” said Yang. “Without WGS, this would have remained an unsolved mystery.”
A new Hybrid Lyme ELISA test shows promise for faster, more accurate early detection of Lyme disease, potentially streamlining lab workflows and improving patient care.
A newly developed serologic test for Lyme disease could significantly improve early detection and streamline the diagnostic process, according to a study published in the Journal of Clinical Microbiology. Researchers say the test, called the Hybrid Lyme ELISA, offers higher sensitivity in the earliest stages of the disease than currently approved methods, potentially reshaping clinical practice.
For laboratory leaders, the emergence of the Hybrid Lyme ELISA marks a potentially pivotal shift in diagnostic testing. With Lyme disease cases approaching half a million annually in the US, labs remain at the forefront of detection and reporting, yet current two-step testing protocols are labor-intensive, time-consuming, and lack sensitivity in early-stage patients. A single-tier, high-sensitivity serologic test could streamline workflows, reduce turnaround times, and enable labs to deliver more actionable results—especially for patients who present with erythema migrans, where current methods fall short.
In a press release on the study, study co-author Gary P. Wormser, MD, New York Medical College said, “This test could potentially change the standard of clinical practice, allowing clinicians to diagnose all manifestations of Lyme disease with a time-saving one-step antibody test.”
Lyme Disease and Current Testing Challenges
Lyme disease, caused by the bacterium Borrelia burgdorferi in the US and related species in Europe, is the most common vector-borne illness in the country, with nearly half a million cases diagnosed each year, according to the Centers for Disease Control and Prevention (CDC). The earliest and most common symptom is a skin rash called erythema migrans, which appears at the site of a tick bite.
If untreated, the infection can spread and cause neurological, cardiac, and joint problems. Early antibiotic treatment is highly effective, but timely diagnosis has remained a challenge.
Since 1995, the CDC has recommended a two-step serology process: an initial antibody screening test, usually an ELISA, followed by a confirmatory test. Originally, the Western Blot was required for confirmation, but in 2019, the CDC allowed a second ELISA instead. While reliable, this sequential approach can delay results.
Neither method is sensitive enough to recommend testing for patients in the erythema migrans stage, leaving early cases often undiagnosed.
A One-Step Approach
Developed by Kephera Diagnostics in collaboration with New York Medical College and the Lyme Disease Biobank, the Hybrid Lyme ELISA employs a novel immunoassay principle. It takes advantage of a unique biological response in which antibodies generated by Borrelia infection can simultaneously bind to two related, though not identical, antigens.
This dual-binding design allows the test to achieve both high sensitivity and specificity in a single step—performance that rivals or exceeds current two-tiered testing.
In the study, the Hybrid ELISA showed sensitivity of more than 90% in patients with erythema migrans, an improvement over the low sensitivity of existing FDA-approved methods. That could make it the first serologic test capable of reliably diagnosing patients in the earliest stage of Lyme disease.
“Current two-tiered serology tests are insensitive in early Lyme disease, missing up to 70% of patients presenting with erythema migrans,” said study co-author Liz Horn, PhD, MBI, principal investigator of the Lyme Disease Biobank, a Bay Area Lyme Foundation program.
Liz Horn, PhD, MBI, principal investigator of the Lyme Disease Biobank noted, “More accurate tests are urgently needed, and the Hybrid ELISA results are very promising.”
Potential to Change Clinical Practice
As an ELISA, the new test can be run manually or on automated high-throughput systems, making it suitable for widespread use in clinical laboratories. Researchers believe it could reduce time, cost, and complexity in diagnosing Lyme disease, while providing earlier answers for patients.
“The publication of this study is an important milestone in our efforts to bring about meaningful improvements in the diagnosis of Lyme disease that will benefit both patients and clinicians,” said Andrew Levin, PhD, chief executive and scientific officer at Kephera Diagnostics. “We are very excited by the results that were achieved by the Hybrid Lyme ELISA in this initial study. Naturally, these findings will have to be corroborated in larger-scale trials, which are currently underway.”
Kephera Diagnostics is now pursuing regulatory approval for the test and plans to make it available through its CLIA-certified laboratory while larger trials continue.
Funding and Collaboration
The Hybrid Lyme ELISA was developed with support from the National Institute of Allergy and Infectious Diseases through a Small Business Innovation Research (SBIR) grant. Collaborators included scientists at New York Medical College and the Lyme Disease Biobank, which provided critical patient samples.
If validated in larger trials and approved, experts say the test could become the first widely available one-step serologic test for Lyme disease, capable of diagnosing patients from the earliest rash stage through later, systemic disease.
