This retrospective, cross-sectional study assessed the burden of HIV in Israel's Maccabi Healthcare Services in 2022. Among 2.6 million individuals assessed (lookback period: > 2 decades), 1973 PWH were identified and age-sex-matched (1:5) to 9,865 randomly selected controls without HIV. We compared sociodemographic, clinical characteristics, and healthcare resource utilization (HRU; Jan-Dec 2022) between people with and without HIV and characterized antiretroviral therapy (ART) treatment patterns among PWH (Jan-Dec 2022). Compared to controls, PWH had a higher lifetime prevalence (since 1988; P < 0.001) of several comorbid conditions, including liver disease, chronic kidney disease, any cancer, and hepatitis B and C infections. Additionally, PWH had a higher rate of anxiety and depression (26.2% vs. 13.5%; P < 0.001). PWH showed higher annual HRU than controls, including ~2-fold higher hospitalizations (≥1 new admissions; P < 0.001) and frequent use of emergency, urgent, primary, specialist, and nursing care (P < 0.05). Among 1907 PWH with ≥1 ART prescription, 78.1% had ≥90% coverage in 2022, although 69.1% experienced ART interruption and 7.2% discontinuation, the latter associated with mental health issues. This study recognizes critical gaps in care that could inform strategies to improve clinical outcomes and resource allocation in health systems for PWH.

BACKGROUND:In 2024, Israel faced a major West Nile virus (WNV) disease outbreak. We aimed to evaluate the efficacy and safety of WNV-neutralizing plasma for hospitalized patients with WNV disease. METHODS:We conducted a randomized, double-blind, placebo-controlled trial. Patients hospitalized with laboratory-confirmed symptomatic WNV disease, 60 years of age or older, or 18 to 59 years of age and immunocompromised, were eligible. Participants were randomly assigned (2:1) to donor-derived WNV-neutralizing plasma or placebo. The primary outcome was a composite of all-cause mortality and any functional deterioration at 30 days, defined as a decrease of more than 5 points from baseline on the Barthel Index (range 0-100; higher scores indicate greater independence) for activities of daily living (i.e., failure to return to pre-illness functional status). Key secondary outcomes included mortality, functional capacity (Barthel Index), and cognitive score (Modified Mini-Mental State Examination; range 0-30; higher scores indicate higher cognitive ability) at 30 and 90 days. RESULTS:Of the 34 participants enrolled, 22 were randomly assigned to the intervention group and 12 to placebo. Median age was 74 (interquartile range, 64-82), 50% were women, and 91% had neuroinvasive disease at enrollment. At 30 days, 11/22 (50%) and 6/12 (50%) patients in the treatment and placebo group, respectively, died or had functional deterioration (risk ratio [RR], 0.96; 95% confidence interval [CI], 0.51-1.79). Two patients (2/22) died in the intervention group, compared with 4 (4/12) in the placebo group (RR, 0.33, 95% CI, 0.09-1.15). The intervention was associated with higher functional capacity (Barthel Index 85 in the intervention group vs. 78 in the placebo group; RR, 1.15; 95% CI, 1.05-1.27), and higher cognitive scores (Mini-Mental score 25 vs. 22; RR, 1.24; 95% CI, 1.04-1.47) at 30 days. One (4.5%) allergic reaction was observed in the intervention group. CONCLUSIONS:In this small, randomized trial of predominantly severe neuroinvasive WNV, treatment with WNV-neutralizing plasma did not reduce the composite outcome of death or functional deterioration at 30 days compared to placebo. It was associated with improved cognitive and functional outcomes. Potential differences in mortality warrant further evaluation. (ClinicalTrials.gov number, NCT06590207.).

OBJECTIVES:To quantify the joint associations of time-varying HIV pre-exposure prophylaxis (PrEP) exposure and bacterial sexually transmitted infections (STIs) testing intensity on detected Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) incidence, and STI-related antibiotic prescribing among men who have sex with men. METHODS:We conducted a retrospective cohort study using electronic health records from Maccabi Healthcare Services, Israel, from January 2019 to June 2024. Men aged ≥21 years with ≥1 rectal CT/NG PCR test were followed in 3-month intervals. PrEP exposure was classified as never, current, recent, or stopped use and lagged by one interval. Testing intensity was defined as the quarterly number of STI testing events. Outcomes were positive PCR results and outpatient prescriptions for doxycycline, azithromycin and ceftriaxone; cephalexin served as a negative control. Adjusted incidence rate ratios (aIRRs) were estimated using mixed-effects negative binomial models including a PrEP-by-testing interaction. RESULTS:Overall, 6943 men contributed 174,516 person-months; 12,573 person-months (7.2%) included ≥1 positive CT/NG PCR result. At annual testing intensity, current PrEP use was associated with higher detected NG and CT incidence than never use (NG aIRR 4.2, 95% CI 3.8-5.3; CT aIRR 5.2, 95% CI 4.0-6.6). These relative differences attenuated with increasing testing intensity. Doxycycline, azithromycin and ceftriaxone prescribing followed similar patterns, whereas cephalexin showed no consistent association with PrEP status or testing intensity. CONCLUSIONS:In PrEP care, testing intensity was strongly associated with detected CT/NG incidence and STI-directed antibiotic prescribing. Risk-stratified testing strategies should be evaluated as part of antimicrobial stewardship in PrEP programmes.

Undocumented and uninsured immigrants living with HIV often face barriers to preventive services, including cancer screening. Evidence is limited regarding how structured screening can be implemented for this population. We evaluated the feasibility, clinical outcomes, and longer-term impact of a cancer screening program delivered through a voluntary HIV-Social clinic in Israel. We conducted a single-center historic–prospective cohort study among undocumented and uninsured immigrants, aged 18 years or above, receiving care at the HIV-Social clinic at Sheba Medical Center. A cancer screening program was established to offer all patients treated in our clinic cancer screening tests, accordingly: cervical cancer screening (Pap smear) to all women, breast imaging to women aged > 45 years, and colonoscopy to adults aged > 50 years. Screening outcomes, follow-up completion, and program costs were recorded. Electronic medical records were reviewed for five years following the program to assess subsequent screening uptake and cancer outcomes. Fifty patients were included (mean age 42 years; 62

BACKGROUND:The high prevalence of asymptomatic Mycoplasma genitalium (MG) infection, combined with limited treatment options due to emerging antimicrobial resistance, raises important questions about the necessity of routine screening and treatment, particularly among men who have sex with men (MSM). This study aimed to determine the prevalence of macrolide and quinolone resistance in MG among HIV-positive MSM and assess its clinical relevance. METHODS:HIV-positive MSM were screened for sexually transmitted infections at the HIV clinic in Sheba Medical Center over a 43-month period, with demographic and clinical parameters recorded upon each visit. A subset of the patients underwent genetic resistance testing for macrolides and quinolones using real-time polymerase chain reaction assays. RESULTS:A total of 317 patients were screened during the study period. MG was detected in 80 (25.2%) patients, covering 150 episodes, at least once during follow-up. In 15 of these episodes (10.0%), patients reported symptoms that might be explained by MG infection. Among the 35 patients who were tested for resistance, 28 (82.3%) harbored macrolide resistance-associated mutations, whereas 11 of 33 (33.3%) exhibited quinolone resistance-associated mutations. CONCLUSIONS:The findings highlight a high prevalence of macrolide-resistant MG infections among HIV-positive MSM, with a notable proportion also exhibiting macrolide quinolone resistance. Given the asymptomatic nature of many MG infections and the increasing challenge of antimicrobial resistance, routine screening for MG in this population may provide limited clinical benefits.

BACKGROUND:Definitions of virological failure and treatment discontinuation for long-acting injectable (LAI) cabotegravir and rilpivirine antiretroviral therapy are inconsistent in clinical practice and observational studies, which complicates interpretation and implementation of findings. The CONSENSUS-LAI study aimed to establish consistent definitions of virological failure and treatment discontinuation to enhance evidence transferability and support optimal clinical outcomes. METHODS:The study had two phases. Phase 1 was an international online survey exploring existing definitions of virological and treatment discontinuation, conducted between April 25 and July 1, 2024. Eligible participants were health-care professionals working in infectious disease or sexual health services who had provided care to at least ten people living with HIV in the past 6 months, had prescribed LAI cabotegravir and rilpivirine in clinical trials or clinical practice, and were able to give informed consent. Participants were recruited via social media and mailing lists of medical specialist societies. Phase 2 was a Delphi process, in which a panel of experts, selected to ensure representation from all six WHO regions, scored leading definitions from phase 1 on a 9-point Likert scale. The proposed definitions were scored according to four validity criteria: clarity, usability in the expert's setting, appropriateness across clinical purposes, and applicability across relevant population groups. Revisions were suggested in iterative rounds until consensus was reached. Consensus was predefined as at least 75% of experts agreeing or strongly agreeing (scores 7-9) with the validity criteria. FINDINGS:386 LAI cabotegravir and rilpivirine prescribers across 28 countries completed the survey, revealing 15 definitions for virological failure on LAI cabotegravir and rilpivirine and nine for treatment discontinuation. 52 experts participated in the Delphi process. Consensus agreement on both definitions was reached after two rounds for all validity criteria. For virological failure, the consensus definition was as follows: (a) viral load 200 copies or more per mL or more on two occasions 2-4 weeks apart, or (b) a single viral load of more than 1000 copies per mL, and/or (c) emergent resistance, in the context of timely injections and prior suppression of less than 200 copies per mL, OR (d) unable to suppress viral load to less than 200 copies per mL on continuous therapy. For treatment discontinuation the consensus definition was as follows: people on LAI cabotegravir and rilpivirine who have missed two consecutive injections and have not taken oral bridging in the interim, irrespective of reason for discontinuation. INTERPRETATION:The consensus definitions provide a foundation for aligning practice and evaluating patient outcomes. Further validation of the viral load threshold for virological failure and the optimal viral load retesting window is required. FUNDING:ViiV Healthcare.

Objective: Assess virological failures, analyze the results of resistance testing (RET), and investigate factors associated with acquired drug resistance mutations (aDRM). Design: A retrospective longitudinal cohort study. Methods: Virological failures (viral load >50 copies/ml) from a cohort of 1130 individuals, diagnosed with HIV-1 in 2010-2018 and followed up until 2020, were included. Demographic, clinical, and virological data were collected. A piecewise exponential additive mixed model was employed to estimate the association of various factors with aDRM. Results: Only 82 individuals had virological failure, 20/82 had multiple virological failures. The majority of virological failures (77%) were men, 48% were Israeli-born,79% were diagnosed in 2010-2014. Only 18% initiated with second-generation integrase-inhibitor (INI) based regimens. Although no baseline differences were identified between those with single and multiple virological failures, the latter had lower CD4(+) levels before first virological failure. NRTI M184IV and INI N155H were identified in more than 10% of the cases. In those with additional failures, INI N155H was more prominent in cases with subtype B compared to those with non-B subtypes (P = 0.039). Diagnoses with CD4(+) cell count less than 200 cells/mu l and AIDS [hazard ratio = 3.46, 95% confidence interval (95% CI): 1.51-7.92, P = 0.003], second-generation INI at the first virological failure (HR = 0.32, 95% CI: 0.11-0.91, P = 0.033), and RET at baseline (hazard ratio = 0.34, 95% CI: 0.13-0.86, P = 0.022) had a significant and persistent relative effect on aDRM. Conclusion: The risk for aDRM is reduced in those who are treated with second-generation INI-based regimens. Diagnosis with low CD4(+) cell counts and AIDS is associated with detection of aDRM.

Abstract Background Despite recent advances in HIV antiretroviral treatment (ART), unmet needs for people with HIV (PWH) remain. The study compared comorbid disease burden and healthcare resource utilization (HRU) among PWH vs those without HIV diagnosis, as well as by ART patterns. Methods A retrospective cross-sectional study used anonymized electronic healthcare data from Israel’s Maccabi Healthcare Services (MHS; >2.7M members) to compare people living with HIV on 31/12/2022 with age-sex-matched controls without HIV diagnosis (1998−2022). PWH had to have ≥1 HIV diagnosis code(s) (1998−2022) and ≥1 dispensed ART prescription (2008−2022). Members enrolled for < 12 months were excluded. Sociodemographic characteristics, lifetime prevalence of comorbidities (exception: anxiety/depression, diagnosed and/or treated in 2022), and HRU (2022) were described. Among PWH dispensed ≥1 ART in 2022, patterns of 12-month percentage of days covered (PDC), discontinuation (≥90-day treatment gap), and interruption (resumed treatment after 6−89-day gap) were described. Results Among 1973 PWH and 9865 controls, mean±SD age was 48.2±11.2 and 48.7±11.2 years, respectively, and 74.4% were male. A lower proportion of PWH (39.6%) vs controls (48.0%) had high residential socioeconomic status. Higher prevalence of comorbidities among PWH vs controls included anxiety/depression (26.2% vs 13.5%), liver disease (20.9% vs 12.5%), chronic kidney disease (14.9% vs 7.3%), cancer (7.2% vs 5.3%), hepatitis C virus (5.6% vs 0.1%), and hepatitis B virus (HBV; 3.4% vs 0.4%). PWH (vs controls) had a median of 10 primary-care visits/year (vs 7) and more frequent hospital admissions (≥1: 11.4% vs 6.6%) and emergency room visits (≥1: 26.5% vs 19.0%). Among PWH treated in 2022 (n=1907), 78.1% had PDC ≥90%. Discontinuation and interruption occurred among 7.2% and 69.1% of treated PWH, respectively. PWH with PDC < 90% (vs ≥90%) had younger age, lower socioeconomic status, higher prevalence of HBV diagnosis, and less frequent physician visits in prior 12 months. Conclusion Despite high ART coverage, PWH in Israel experience an excess burden of comorbidities and HRU compared to age-sex-matched controls without HIV. Further research is needed to address the excess disease burden and better define treatment/care gaps among PWH. Disclosures Sivan Gazit, MD, AbbVie: The relationship is not personal, AbbVie funded the current study, funding to the research institution where I work Arlene Nugent, MSc, Abbvie: Stocks/Bonds (Public Company) Liza Bronner Murrison, PhD, AbbVie: Stocks/Bonds (Public Company) Ana Gabriela Pires dos Santos, PhD, ABBVIE: Stocks/Bonds (Public Company)

In Israel, irregular migrants (IMs) living with human immunodeficiency virus (HIV-1) that have no access to regular health insurance are provided with HIV-1-related health coverage under a public–private partnership (PPP) program initiated by the Ministry of Health in 2014. Here we characterized IMs referred to the PPP in 2019–2024 and used a linear mixed-effects model to follow up their CD4 and HIV-1 viral load (VL) counts for a median period of 16 months. Subtypes, resistance mutations and phylogenetic relationships were determined in all cases with viral failure and in selected cases with available blood remains. A total of 231 of 238 referred to the PPP initiated antiretroviral treatment (ART) with nucleoside reverse transcriptase inhibitors (NRTIs) and either non-nucleoside reverse transcriptase inhibitors (NNRTIs, 61.5%, 142/231) or protease inhibitors (PIs, 38.5%, 89/231). Irrespective of the treatment regimen, all these individuals increased their CD4 and decreased their VL trajectories over time (p < 0.001). However, mixed model analysis revealed two classes of CD4 trajectory patterns. Comparison between these two patterns revealed that Class-1 individuals started with lower initial CD4 counts compared to Class-2 individuals (median of 115 cells/mm3, IQR 70–171 compared to median of 312 cells/mm3, IQR 104–510, p < 0.001) and experienced slower recovery compared to Class-2. Most Class-1 individuals originated from Africa (78% vs. 52%, p = 0.016). Treatment failure was observed in 5.6% of all individuals receiving treatment under the program. Sequencing analysis, enabled in 23% of the treated cohort, revealed that the reverse transcriptases (RT) M184V (13%) and K103N (7.4%) were the most prevalent mutations. Conclusively, while treatment was not consistent with current recommendations for first-line therapy, the virological and immunological response of most patients was favorable and the prevalence of cases with resistance mutations was not higher than that identified in people living with HIV-1 who are covered by the national health insurance. Despite the limitations of the PPP, this program may provide a unique therapeutic opportunity for IMs with HIV-1.

Since May 2024, Israel has been experiencing a large West Nile virus (WNV) outbreak. We aimed to compare the clinical characteristics and outcomes of hospitalized cases to previous years and identify predictors of poor outcomes. A retrospective study. We compared WNV infection cases hospitalized during the 2024 outbreak (from 29 May to 29 July) to cases hospitalized during 2018–2023. For the entire cohort, risk factors for poor outcomes were investigated using multivariable analyses. The primary outcomes were death and a composite outcome of 30-day all-cause mortality, prolonged hospitalization (≥ 28 days), or discharge to an institution. We included 134 patients, 103 admitted during 2024 and 31 during 2018–2023. The majority (109/134, 81

WHAT IS THIS SUMMARY ABOUT?:This is a summary of an article about an ongoing study called the BICSTaR study.The BICSTaR study includes people with HIV (human immunodeficiency virus) who are taking a medicine called bictegravir/emtricitabine/tenofovir alafenamide (shortened to B/F/TAF). B/F/TAF is a single tablet that contains 3 different drugs for the treatment of HIV. The drugs work together to reduce the levels of HIV so that the virus can no longer be detected by a blood test.People taking part in the study are adults with HIV living in Europe, Canada, Israel, Japan, South Korea, Singapore and Taiwan. People take 1 tablet of B/F/TAF once a day. They are either taking B/F/TAF as their first treatment for HIV, or they have switched to B/F/TAF from another HIV treatment.Researchers looked at how well B/F/TAF worked and how safe it was in people who took B/F/TAF for a year. WHAT ARE THE KEY TAKEAWAYS?:Researchers found that B/F/TAF worked well in almost all people in the study by reducing levels of HIV in the blood. The virus could not be found in the blood of more than 9 out of 10 (94%) people who were taking B/F/TAF as their first HIV medicine and more than 9 out of 10 people (97%) who had taken another HIV medicine before starting B/F/TAF. This is known as having an 'undetectable viral load' and is a major goal for HIV treatment success. Researchers did not find any evidence of HIV developing resistance to B/F/TAF, which might stop B/F/TAF from working properly.Around 1 out of 10 people (13%) had side effects (any unwanted sign or symptom that people have when taking a medicine that researchers think might be caused by the medicine) that might have been caused by B/F/TAF. Most of these side effects were not classified as serious. Less than 1 out of 100 (0.1%) people had serious side effects that might have been caused by B/F/TAF. Only 6 out of 100 people stopped taking B/F/TAF due to side effects caused by B/F/TAF. As a result, more than 9 out of 10 people (95%) took B/F/TAF for at least 1 year. WHAT WERE THE MAIN CONCLUSIONS REPORTED BY THE RESEARCHERS?:B/F/TAF worked well in people with HIV in this study. Most people (around 9 out of 10) did not have any side effects.

Diabetes mellitus (DM) is more common among people living with human immunodeficiency virus (PLWH) compared with healthy individuals. In a prospective multicenter study (N = 248), we identified normoglycemic (48.7%), prediabetic (44.4%), and diabetic (6.9%) PLWH. Glycosylated hemoglobin (HbA1c) and fasting blood glucose (FBG) sensitivity in defining dysglycemia was 96.8%, while addition of oral glucose tolerance test led to reclassification of only 4 patients. Inclusion of 93 additional PLWH with known DM enabled identification of multiple independent predictors of dysglycemia or diabetes: older age, higher body mass index, Ethiopian origin, HIV duration, lower integrase inhibitor exposure, and advanced disease at diagnosis. Shotgun metagenomic microbiome analysis revealed 4 species that were significantly expanded with hyperglycemia/hyperinsulinemia, and 2 species that were differentially more prevalent in prediabetic/diabetic PLWH. Collectively, we uncover multiple potential host and microbiome predictors of altered glycemic status in PLWH, while demonstrating that FBG and HbA1c likely suffice for diabetes screening. These potential diabetic predictors merit future prospective validation.

BACKGROUND:Real-world evidence is an essential component of evidence-based medicine. The aim of the BICSTaR (BICtegravir Single Tablet Regimen) study is to assess effectiveness and safety of bictegravir/emtricitabine/tenofovir alafenamide (B/F/TAF) in antiretroviral treatment-naïve (TN) and treatment-experienced (TE) people with HIV. METHODS:BICSTaR is a prospective, observational cohort study. Participants (≥18 years) are being followed for 24 months. A pooled analysis is presented at 12 months, with the primary endpoint of effectiveness (HIV-1 RNA <50 copies/mL) and secondary endpoints of safety and tolerability (as per protocol). An exploration of patient-reported outcome measures using standardized questionnaires is included. RESULTS:Between June 2018 and May 2021, 1552 people with HIV were enrolled across 12 countries. The analysed population comprised 1509 individuals (279 TN, 1230 TE); most were white (76%), male (84%) and had one or more comorbid conditions (68%). Median age was 47 years. After 12 months of B/F/TAF treatment, HIV-1 RNA was <50 copies/mL in 94% (221/236) of TN participants and 97% (977/1008) of TE participants. Median CD4 cell count increased by 214 cells/μL (p < 0.001) in TN participants and 13 cells/μL (p = 0.014) in TE participants; median CD4/CD8 ratios increased by 0.30 and 0.03, respectively (both p < 0.001). Persistence was high at 12 months (TN, 97%; TE, 95%). No resistance to B/F/TAF emerged. Study drug-related adverse events occurred in 13% of participants through 12 months, leading to B/F/TAF discontinuation in 6%. CONCLUSIONS:The findings of this study provide robust real-world evidence to support the broad use of B/F/TAF in both TN and TE people with HIV.

Despite the progress in contemporary antiretroviral therapy (ART) and the continuous changes in treatment guidelines, virological failure (VF) is still an ongoing concern. The goal of this study was to assess factors related to VF after first-line ART. A longitudinal cohort retrospective study of individuals on first-line ART diagnosed with HIV-1 in 2010–2018 and followed-up for a median of two years was conducted. Demographics, baseline and longitudinal CD4 counts, treatment regimens, adherence and VF were recorded. The Cox proportional hazards regression and mixed models were used. A cohort of 1130 patients were included. Overall, 80% were males and 62% were Israeli-born individuals. Compared to individuals diagnosed in 2010–2014, when treatment was initiated according to CD4 levels, those diagnosed in 2015–2018 were older and had lower baseline CD4 counts. VF was recorded in 66 (5.8%) patients. Diagnosis with CD4 <200 cells/mmᶟ with AIDS-defining conditions (HR = 2.75, 95%CI:1.52–4.97, p < 0.001) and non-integrase strand transfer inhibitor regimens (non-INSTI, HR = 1.80, 95%CI:1.01–3.24, p = 0.047) increased VF risk. No impact of baseline resistance was observed. We concluded that the early detection of HIV-1 infection and usage of INSTI-based regimens are recommended to reduce VF.

PURPOSE OF REVIEW:Persons living with HIV (PLWH) may have a moderately increased risk of morbidity and mortality from COVID-19 infection, especially if viral load is not controlled and if they are immunosuppressed. Vaccination against SARS-CoV-2 is the most effective measure to prevent morbidity and mortality. However, individuals with HIV/AIDS may have less protection after vaccination. The purpose of this review is to summarize some of the recent studies focused on examining the safety, immunogenicity and effectiveness of anti-SARS-CoV-2 vaccines.RECENT FINDINGS:The safety of all anti-SARS-CoV-2 vaccines among PLWH is not different from the safety of these vaccines among HIV-negative individuals and is acceptable. PLWH with viral suppression and immune reconstitution (CD4 + cell count > 350 cells/μl) may reach almost same immunogenicity such as people without HIV albeit antibody levels and neutralization may decline more rapidly than in people without HIV. PLWH with viremia or immunosuppressed, especially AIDS, have less immunogenicity.SUMMARY:Full vaccination against SARS-CoV-2 is a well tolerated and efficient way to prevent mortality and morbidity from COVID-19 among PLWH and AIDS patients. It is very important to follow recommended booster vaccination for a continuous and prompt immunogenicity.

In this report, we describe the first national scale multi-laboratory evaluation of monkeypox virus (MPXV) DNA commercial PCR kits. The objective of this study was to evaluate 2 kits by different diagnostic laboratories across Israel. Ten standardized samples were tested simultaneously using the Novaplex (15 laboratories) and Bio-Speedy (seven laboratories) kits. An in-house assay based on previously published reactions was used as reference. Comparison of the results showed high intra-assay agreement between laboratories, with small variations for most samples. The in-house assay had an analytical detection limit of less than 10 copies per reaction. While the 2 commercial kits were able to detect specimens with low viral loads similarly to the in-house assay, significant differences were observed, in the Cq values and relative fluorescence (RF), between the assays. The RF signal of the in-house and Bio-Speedy assays ranged between 5,000 and 10,000 RFU, while the signal in the Novaplex assay was less than 600 RFU. Due to the kit measurement protocol, the Cq values of the Bio-Speedy kit were 5 to 7.5 cycles lower than those of the in-house assay. On the contrary, the Cq values of the Novaplex kit were significantly higher than those of the in-house assay, with differences of 3 to 5 cycles per sample. Our results suggest that while all assays were similar in their overall sensitivity, direct comparison of Cq values between them may be misleading. To our knowledge, this is the first methodical evaluation of commercial MPX test kits. We therefore anticipate that this study would help diagnostic laboratories in choosing a specific MPX detection assay. IMPORTANCE To the best of our knowledge, this study is the first methodical evaluation of commercial kits designed for Monkeypox virus detection. This was done by performing the same tests using the same sample set in multiple laboratories, simultaneously, on a national scale. It therefore provides important and unique information on the performance of such kits and provides a guideline for choosing the assay of choice for monkeypox virus diagnosis in a standard diagnostic laboratory. It also demonstrates potential complications when trying to compare the results of different assays, even when testing exactly the same samples, under identical conditions.