Rapid Detection Test Research Articles

Molecular Rapid Test for Identification of Tuna Species.

Tuna is an excellent food product, relatively low in calories, that is recommended for a balanced diet. The continuously increasing demand, especially for bluefin-tuna-based food preparations, and its relatively high market price make adulteration by intentionally mixing with other lower-priced tunas more prospective. The development of rapid methods to detect tuna adulteration is a great challenge in food analytical science. We have thus developed a simple, fast, and low-cost molecular rapid test for the visual detection of tuna adulteration. It is the first sensor developed for tuna authenticity testing. The three species studied were Thunnus thynnus (BFT), Thunnus albacares, and Katsuwonus pelamis. DNA was isolated from fresh and heat-treated cooked fish samples followed by PCR. The PCR products were hybridized (10 min) to specific probes and applied to the rapid sensing device. The signal was observed visually in 10-15 min using gold nanoparticle reporters. The method was evaluated employing binary mixtures of PCR products from fresh tissues and mixtures of DNA isolates from heat-treated tissues (canned products) at adulteration percentages of 1-100%. The results showed that the method was reproducible and specific for each tuna species. As low as 1% of tuna adulteration was detected with the naked eye.

Development of Enzyme-Linked Immunosorbent and Immunochromatography Assays for Diagnosing Nosema ceranae Infection in Honey Bees.

Nosema ceranae (N. ceranae) infection is prevalent globally, causing a decline in bee populations and significant economic losses to apiarists. Although several methods have been proposed for diagnosing Nosema infections, limitations in these methods have hindered their broad applications. Therefore, this current study aimed to develop a specialized method for diagnosing Nosema infections. To achieve this, a sandwich enzyme-linked immunosorbent assay (ELISA) and immunochromatography assay (ICG) were developed, and their effectiveness in screening and diagnosing Nosema infection was assessed. In sandwich ELISA, the combination of the monoclonal antibodies (mAb) 19B2 and biotinylated-19B2 exhibited stronger binding affinity to the antigen than did other combinations of mAbs that were tested. Furthermore, the antigen detection limit achieved with the sandwich ELISA surpassed that previously reported with Western blotting. The ICG was designed using the same antibody combination as that used in sandwich ELISA; however, the assay exhibited a lower diagnostic ability for Nosema infection than the ELISA. The diagnostic models developed in this study offer practical applications for conducting rapid nosemosis detection tests. These innovative techniques will help to improve the timely identification and management of nosemosis.

Evaluation of the Xpert Xpress GBS test for rapid detection of group B Streptococcus in pregnant women.

This was the first study evaluating the performance of the Xpert Xpress group B Streptococcus (GBS) test using rectovaginal swabs from Chinese pregnant women. Compared to the other three assays, the Xpert Xpress GBS test demonstrated high sensitivity and specificity when screening 939 pregnant women for GBS in rectovaginal specimens. Additionally, its reduced time to obtain results makes it valuable for the rapid detection of GBS.

A Short Communication: An Imported Case of <i>Plasmodium falciparum</i> Malaria Infection from Ivory Coast leave to Iran

Malaria is the most common cause of fever in travellers returning from malaria-endemic areas. Imported malaria can hinder the progress of elimination programs. In the present study, we describe an imported malaria case in a 45-year-old Iranian man from Ivory Coast who had returned to Iran after taking annual leave. He was hospitalized in Kamkar-Arabnia Hospital, Qom, central Iran, in January 2023. Symptoms were fever, chills, sweating, cough, myalgia, sweating, anorexia, diarrhoea, and headache. Rapid detection test and blood smears were positive for Plasmodium falciparum gametocyte. The patient was treated with an antimalarial drug and discharged from the hospital with normal vital signs. Health education is recommended for people before leaving and screening after returning from travel. In addition, using RDT tests for screening people who return to the country is suggested to health staff and organizations.

CER818: A Highly Specific and Sensitive HPV L1 High-Risk Serological Lateral Flow Rapid Test for Early Detection of Cervical Cancer and Its Precursor Lesions.

Objective: The objective of the study is to validate a new human papillomavirus (HPV) L1 high-risk specific serological assay in a case-control study. Methods: Serum samples of 138 patients (cervical intraepithelial neoplasia (CIN) 1, 2, and 3 and cervical cancer), 21 vaccinees, and 246 female controls were tested for the presence of HPV L1 high-risk specific antibodies. Results: HPV L1 high-risk antibodies were detected in 100% of the CIN1 and 2, 86.6% of the CIN3 and 82.4% of the cervical cancer cases, 100% of the vaccinees, and 3.9% of the female controls. Area under the curve (AUC) was calculated with 0.91 for controls versus CIN2+, 0.923 for controls versus CIN1+, and 0.968 for controls versus CIN1/2. Conclusion: The HPV L1 high-risk specific serological lateral flow rapid test shows promising data in the field of early detection of HPV high-risk induced cervical cancer and its precursor lesions. This easy-to-use, robust, and affordable approach could offer a chance to reach women in low- or middle-income countries (LMICs) that could not be reached by HPV molecular testing-based cervical cancer screening programs.

Is near-infrared spectroscopy a promising predictor for early intracranial hemorrhage diagnosis in the Emergency Department?

Intracranial hemorrhage (ICH) is a serious medical condition that can lead to significant morbidity and mortality if not diagnosed and treated promptly. Early detection and treatment are essential for improving the outcome in patients with ICH. Near-infrared spectroscopy (NIRS) is a non-invasive imaging technique that has been used to detect changes in brain tissue oxygenation and blood flow in various conditions. The aim of this study was to investigate the predictive potential of NIRS for early diagnosis of ICH in patients presenting to the Emergency Department (ED) triage with headache. A total of 378 patients were included in the study. According to the final diagnosis of the patients, 4 groups were formed: migraine, tension-cluster headache, intracranial hemorrhage and intracranial mass, and control group. Cerebral NIRS values "rSO2" were measured at the first professional medical contact with the patient. The right and left rSO2 (RrSO2, LrSO2) were significantly lower and the rSO2 difference was significantly higher in the intracranial hemorrhage group compared to all other patient groups (P<0.001). The cut-off values determined in the receiver operating characteristics (ROC) analysis were RrSO2 ≤67, LrSO2 ≤67, and ΔrSO2 ≥9. This study found that a difference of more than 9 in cerebral right-left NIRS values can be a non-invasive, easy-to-administer, rapid, and reliable diagnostic test for early detection of intracranial bleeding. NIRS holds promise as an objective method in ED triage for patients with intracranial hemorrhage. However, further research is needed to fully understand the potential benefits and limitations of this method.

Effect of hepatitis B vaccination on HBV-infection among school children in Yaounde; ten years after the introduction of HBV vaccine into routine Immunization Program in Cameroon.

since the introduction of the anti-HBV vaccine into the Expanded Program on Immunization (EPI) in 2005 in Cameroon, vaccination coverage has reached 99.0%. This coverage would indicate an increase in the number of children immune to Hepatitis B Virus (HBV) and a decrease in susceptibility to HBV-infection. This study was conducted to evaluate the effect of the HBV vaccine on pediatric HBV-infection in Yaounde, Cameroon. this school-based cross-sectional study was conducted from February to May 2016 among 180 children from Nkomo public school. The study population was stratified into two groups: vaccinated (n=95) versus (vs) unvaccinated (n=85). Screening for HBV biomarkers was done using a rapid panel test for detection (HBsAg, HBeAg and anti-HBc) and anti-HBs titer using enzyme linked immunosorbent assay (ELISA). Statistical analyses were done using SPSS v. 22 with p < 0.05 considered significant. the mean age was 9.65 years. HBsAg (p=0.019) and anti-HBc (p=0.001) rates were detected in children aged ≥10 years and children aged < 10 years (95.95% [71/74]) were vaccinated vs 22.64% (24/106) for those aged ≥10 years (OR: 80.86; 95% CI: 23.36%-279.87%, p < 0.0001). According to anti-HBV vaccination status, HBsAg rate varied from [9.41% (8/85) to 1.05% (1/95), p=0.025], HBeAg rate varied from [2.35% (2/85) to 0% (0/95), p= 0.42] and anti-HBc rate ranged from [12.94% (11/85) to 2.10% (2/95), p= 0.011]. despite the variability of the anti-HBs titer, vaccination against HBV has a positive effect on the reduction of HBV-infection in children in tropical settings such as Cameroon.

The use of colour and rapid flicker sensitivity tests for early detection and monitoring of vision changes in diabetes

Aims/Purpose: There is increasing evidence of impaired visual function prior to the presence of diabetic retinopathy (DR), yet current diagnostic criteria and treatment onset rely mostly on retinal vascular changes. In this study we measured changes in chromatic and rapid flicker sensitivity in subjects with prediabetes and diabetes to establish the value of such tests as a potential biomarker in the detecting early diabetes.Methods: Three groups of subjects were recruited: 1) normal controls, 2) subjects with prediabetes (HbA1c between 42‐47 mmol/mol), and 3) diabetics (with or without DR). Monocular cone‐ and rod‐enhanced flicker modulation thresholds (FMTs) were measured at the point of regard and in four parafoveal (5°) locations using the Flicker‐Plus test. Monocular red/green (RG) and yellow/blue (YB) colour thresholds were also measured at the fovea using the Colour Assessment and Diagnosis (CAD) test.Results: Both the RG and YB thresholds, as well as rod‐ and cone‐mediated flicker thresholds were significantly higher in both diabetic and prediabetic groups (when compared with healthy controls). When examined individually, ~50% and ~ 63% of the prediabetics and ~ 58% and ~ 70% of the diabetics revealed abnormal RG and YB thresholds, respectively (when compared with the upper normal limits for the corresponding age). HBA1c levels also revealed a positive trend with increasing RG and YB thresholds in the prediabetic group.Conclusions: In agreement with previous findings, both colour and FMTs are affected in patients diagnosed with diabetes. Unexpectedly, those deemed prediabetic that do not meet the clinical criteria for diabetes exhibit considerable loss of both colour and flicker sensitivity. The results suggest that colour and rapid flicker tests can provide useful information when screening for diabetes.

A Clinical Evaluation of the HPV Antigen Rapid Test for Early Detection of Cervical Cancer

A Clinical Evaluation of the HPV Antigen Rapid Test for Early Detection of Cervical Cancer

Evaluation of the Fecal Antigen Rapid Test for Early Detection of Helicobacter Pylori Infection

Evaluation of the Fecal Antigen Rapid Test for Early Detection of Helicobacter Pylori Infection

Development and Application of a Rapid Test for Detection of Bovine Viral Diarrhea Virus-specific Antibodies

Development and Application of a Rapid Test for Detection of Bovine Viral Diarrhea Virus-specific Antibodies

Performance Evaluation of Multi-Drug Rapid Test for Rapid Detection of Multiple Drugs and Drug Metabolites in Human Blood Samples

Objective: The Multi-Drug Rapid Test Cassette, developed by Hangzhou AllTest Biotech Co., Ltd, is a rapid chromatographic immunoassay designed for the qualitative detection of multiple drugs and drug metabolites in human whole blood, serum, or plasma. It aims to simultaneously detect various abused substances, including Amphetamine (AMP), Barbiturates (BAR), Benzodiazepines (BZO), Buprenorphine (BUP), Cocaine (COC), Cannabis (THC), Methadone (MTD), Methamphetamine (MET), 3,4-Methylenedioxymethamphetamine (MDMA), Morphine (MOP/OPI), Propoxyphene (PPX), Tricyclic antidepressants (TCA), Oxycodone (OXY), 2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), Cotinine (COT), Tramadol (TML), Fentanyl (FYL), 3,4-Methylenedioxy pyrovalerone (MDPV), Synthetic cannabinoids(K2), Phencyclidine(PCP), Ketamine(KET), Lysergic acid diethylamide(LSD), 3,4-Methylenedioxyamphetamine(MDA), Acetaminophen(ACE), Ketamine(CAT), 6-Monoacetylmorphine (6-MAM), Zolpidem (ZOL), AB-PINACA (ABP/K3). Therefore, the present research was done to evaluate the diagnostic performance of this test. Material and Method: In this study, the accuracy, precision, and sensitivity of the drug testing methods were assessed. Approximately 100 specimens of each drug, obtained from previous drug screening tests, were tested. The results were compared to confirmatory analysis using GC/MS, and the accuracy of the testing was determined based on the comparative classification results. The evaluation of precision included measuring the consistency of measurements within a single run, between different runs, and among different operators, using the AllTest multi-drug detection tool across three different hospitals and three different batches. To evaluate the sensitivity of the detection method, various concentrations of drugs were added to a pool of whole blood, serum, or plasma without any drugs. The positive and negative results obtained from the testing were analyzed to assess the method’s ability to detect drugs at different concentrations. Results: In this study, we evaluated the performance of the Multi-Drug Rapid Test Cassette developed by Hangzhou AllTest Biotech Co., Ltd. The results demonstrated excellent performance of the test in terms of accuracy, sensitivity, and precision. Conclusion: The Multi-Drug Rapid Test Cassette developed by Hangzhou AllTest Biotech Co., Ltd exhibited reliable detection of multiple drugs and drug metabolites, providing trustworthy results for drug abuse screening. These findings have significant implications for drug control and monitoring, as well as supporting the broad application of this test in clinical and legal settings.

Ultrasensitive biosensing platform for Mycobacterium tuberculosis detection based on functionalized graphene devices.

Tuberculosis (TB) has high morbidity as a chronic infectious disease transmitted mainly through the respiratory tract. However, the conventional diagnosis methods for TB are time-consuming and require specialists, making the diagnosis of TB with point-of-care (POC) detection difficult. Here, we developed a graphene-based field-effect transistor (GFET) biosensor for detecting the MPT64 protein of Mycobacterium tuberculosis with high sensitivity as a POC detection platform for TB. For effective conjugation of antibodies, the graphene channels of the GFET were functionalized by immobilizing 1,5-diaminonaphthalene (1,5-DAN) and glutaraldehyde linker molecules onto the graphene surface. The successful immobilization of linker molecules with spatial uniformity on the graphene surface and subsequent antibody conjugation were confirmed by Raman spectroscopy and X-ray photoelectron spectroscopy. The GFET functionalized with MPT64 antibodies showed MPT64 detection with a detection limit of 1fg/mL in real-time, indicating that the GFET biosensor is highly sensitive. Compared to rapid detection tests (RDT) and enzyme-linked immunosorbent assays, the GFET biosensor platform developed in this study showed much higher sensitivity but much smaller dynamic range. Due to its high sensitivity, the GFET biosensor platform can bridge the gap between time-consuming molecular diagnostics and low-sensitivity RDT, potentially aiding in early detection or management of relapses in infectious diseases.

Food safety and quality training: The use of a lateral flow immunochromatographic strip test for rapid detection of pesticide residue on leafy vegetables

Ensuring the safety and quality of leafy vegetables, especially by controlling any traces of pesticides, is crucial for maintaining human health. This international community service program was designed to expand the vocational knowledge and skills of the people of Timor-Leste in using lateral flow immunochromatographic strip tests for quickly detecting pesticide residue on leafy vegetables. The program was organized by UAD in partnership with PCIM Timor-Leste and the An-Nur Islamic Foundation. The program included several stages, including preparation, coordination, consolidation, implementation, monitoring, and evaluation. The participants’ knowledge of food safety and food hygiene increased significantly from 52% prior to training to 96% after the training. Moreover, the ability to detect pesticide contamination in vegetables using LFI technology improved by 84%, rising from just 8% before training to 92% after training. All participants expressed interest in the UAD in Timor-Leste's future continuation of this training program.

Molecular typing of canine parvovirus type 2 by VP2 gene sequencing and restriction fragment length polymorphism in affected dogs from Egypt.

Canine parvovirus-2 (CPV-2) is one of the most common infectious diseases in dogs characterized by severe gastroenteritis, vomiting, and bloody diarrhea. Little information is available about this topic in Egypt, particularly in the Delta region. This study reports the prevalence and molecular analysis of CPV-2 variants collected from El-Gharbia and Kafrelsheikh governorates in the Delta of Egypt. In this study, 320 rectal swabs were collected from infected domestic dogs from two districts in delta Egypt. The samples were investigated by rapid immunochromatographic test and polymerase chain reaction for detection the prevalence of CPV-2 variants. The genetic characterization was performed using restriction fragment length polymorphism (RFLP) analysis and partial VP2 gene sequence. The viral antigen was detected in (264/320, 82.5%) of samples by IC test, while PCR was found more sensitive by detecting (272/320, 85%) positive samples. The RFLP technique using MboII restriction enzyme was successfully used for the differentiation of CPV-2c antigenic variants from CPV-2a/2b strains. Interestingly, the molecular and phylogenetic analysis revealed that both CPV-2a and CPV-2c are circulating in the study area. Deduced amino acid sequence analysis showed changes at residue (N426E) and residue (T440A).: Our results indicated that CPV-2 is prevalent among dogs in Egypt, and therefore further molecular and epidemiological studies of CPV-2 are warranted.

Antibody-labelled gold nanoparticles synthesized by laser ablation to detect SARS-CoV-2 antigen spike.

Rapid detection test via lateral flow immunoassay (LFIA) is employed as an alternate method to detect Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection. Gold nanoparticles (AuNPs), a vital component of LFIA, can be synthesized by laser ablation technique. This intense laser radiation may result in monodisperse gold nanoclusters, which are impurity-free and demonstrate innovative biocompatible surface chemistry. In this current research, laser-ablated AuNPs are produced and coupled with an anti-spike SARS-CoV-2 monoclonal antibody (mAb) generated in our prior study. The AuNPs from 30,000 shots of laser ablation exhibited a robust red color with a maximum absorbance peak at 520 nm. The performance of AuNPs-mAb conjugates as a signal reporter was then evaluated in half-stick LFIA. The size distribution of AuNPs shows a relatively monodisperse and unimodal distribution with average particle diameters of 44.77 nm and a surface potential of -38.5 mV. The purified anti-spike mAb SARS-CoV-2 yielded two protein bands, representing the mAb heavy chain at 55 kDa and its light chain at 25 kDa. The immobilization of anti-spike mAb onto the surface of AuNPs revealed that 25 g/ml of mAb at phosphate buffer pH 9 was required to stabilize the AuNPs. The functional test of this conjugate was performed using dipstick LFIA, and the result shows that the AuNPs-mAb conjugates could successfully detect commercial spike antigen of SARS-CoV-2 at 10 ng level. In this study, laser-ablated AuNPs were functionalized with anti-spike mAb SARS-CoV-2 and successfully used as a signal reporter in half-stick LFIA for detecting antigen spike SARS-CoV-2.

Rapid immunochromatographic test: An evolving tool for diagnosis of scrub typhus

Background: Scrub typhus is prevalent in many districts of South Bengal throughout the year where an average temperature of 20–35°C, which contributes to the spread of Leptotrombidium deliense. However, its diagnosis remains complicated by the lack of readily available and validated assays, the non-specificity of clinical symptoms on admission, and even non-availability of the pathognomonic eschar in most of the cases. Aims and Objectives: This study was carried out to evaluate the rapid immunochromatographic test (RICT) for early detection of scrub typhus for using it as an early diagnostic tool at the field level. Materials and Methods: This cross-sectional study in which 181 serum samples from clinically suspected cases (after excluding dengue, malaria, Japanese encephalitis, and typhoid fever) collected over 13 months were processed for the detection of immunoglobulin M (IgM) antibodies for scrub typhus by enzyme-linked immunosorbent assay (ELISA) and rapid test. Results: Considering IgM ELISA for scrub typhus as the gold standard, the sensitivity, specificity, positive predictive value, and negative predictive value for RICT were found to be 100%, 86.87%, 50%, and 100%, respectively. Conclusion: RICT is a simple, rapid, and reliable assay for diagnosis of scrub typhus, capable of providing accurate results quickly and is highly suitable for field deployment in remote areas with limited medical support.

Establishment of a Colloidal Gold Immunochromatographic Method for the Detection of Cypermethrin in Tobacco.

In this study, the establishment of a colloidal gold immunochromatographic method for the detection of cypermethrin in tobacco was achieved by using colloidal gold immunochromatography: strong specificity and high sensitivity of cypermethrin semi-antigens and encapsulants were prepared during the study. The best colloidal gold solution was prepared by spectrophotometer and transmission electron microscope screening; the preparation process of gold-labeled antibodies was optimized, and finally the product of colloidal gold rapid detection test strips for cypermethrin was developed. The results of technical parameters and detection indexes showed that the detection limit of cypermethrin in tobacco was 1 mg/kg, and there was no cross-reaction with bifenthrin, cypermethrin, cyfluthrin and phenothrin, and the detection results of 30 tobacco samples were consistent with those of gas chromatography.

Accuracy of a new rapid diagnostic test for urinary antigen detection and assessment of drug treatment in opisthorchiasis

BackgroundScreening for opisthorchiasis, a parasitic worm infection affecting many millions of people in Southeast Asia, has traditionally relied on faecal egg examination such as the formalin-ethyl acetate concentration technique (FECT) and Kato-Katz method. Although the urinary enzyme-linked immunosorbent assay (ELISA) has been used more recently, we developed a urinary antigen-based rapid diagnostic test (RDT) to simplify diagnosis and as a point-of-care testing (POCT) and field applications for surveillance and control of opisthorchiasis.MethodsA urinary Opisthorchis viverrini (OV)-RDT was developed using immunochromatographic methodology with a specific monoclonal antibody against OV. The diagnostic performance of the urinary OV-RDT was compared to that of quantitative faecal FECT and urinary antigen ELISA (n = 493). Cross-reactivities of urinary OV-RDT with other helminthiases coexisted with O. viverrini were determined (n = 96). A field trial in the application of urinary OV-RDT was compared with urinary antigen ELISA at baseline screening and assessment of drug treatment outcomes in opisthorchiasis (n = 1629). The McNemar chi-square, Kruskal–Wallis and Cohen’s kappa coefficient (κ-value) tests were used for statistical analyses.ResultsUrinary OV-RDT had sensitivity of 94.2% and specificity of 93.2%, compared to faecal FECT. Urinary OV-RDT had high diagnostic agreement (Kappa = 0.842–0.874, P < 0.001) and quantitative correlation with urinary antigen ELISA (Kruskal–Wallis tests = 316.2, P < 0.0001) and faecal FECT (Kruskal–Wallis tests = 362.3, P < 0.0001). The positive rates by OV-RDT, ELISA and FECT were 48.9%, 52.5% and 49.3%, respectively. Cross-reactions of urinary OV-RDT with other helminthiases were few (2%). Field trials of urinary OV-RDT yielded comparable prevalence of O. viverrini between urinary OV-RDT (53.2%) and urinary antigen ELISA (54.0%). OV screening showed high diagnostic agreement (kappa > 0.8, P < 0.0001) between urinary OV-RDT and urinary antigen ELISA. The cure rates of opisthorchiasis at 1 month post-praziquantel treatment determined by urinary OV-RDT (86.6%) and urinary antigen ELISA (80.5%) were similar (P > 0.05).ConclusionsThe urinary OV-RDT test has high potential as a new tool for screening and evaluating treatment outcomes in opisthorchiasis. The ease of sample collection and simplicity of urinary OV-RDT may facilitate mass screening, control and elimination of opisthorchiasis, thereby contributing to a reduction in the disease burden in Southeast Asia.Graphical

Changing trend analysis on early detection of rifampicin resistant tuberculosis patients in southwestern area of China, 2016-2020.

There were no data about prevention and control status of RR-TB in a poor area with high burden of TB in China. In order to develop evidence-based RR-TB response strategies and improve enrollment of RR-TB patients in Yunnan province, China, this study was aimed at analyzing the changing trends in the detection and enrollment of RR-TB patients and examining the factors that may have implication on enrollment in treatment. Data, which includes demographics, screening and testing, and treatment enrollment, was collected from the TB Management Information System. Retrospective data analysis and factors analysis were applied. Descriptive statistics, Chi-square test, Rank sum test and logistic regression analysis were used. From 2016 and 2018, the province had been challenged by low levels of screening, detection and enrollment of RR-TB. During the period between 2019 and 2020, a comprehensive model of RR-TB prevention and control was established in Yunnan, characterized by a robust patient-centered approach for RR-TB care and multiple, targeted interventions through the cascade of care from detection to treatment. In 2020, 93.8% of the bacteriologically positive TB patients were screened for RR-TB, which had been significantly increased by 146.9% from 38.0% in 2016. The interval from initial consultation at RR-TB facility to diagnosis (inter-quartile range) was reduced from 29.5 (1-118) days in 2016 to 0 (0-7) days in 2020. Despite the increasing rates of enrollment of RR-TB patients over the years, non-enrollment of those detected was still high (32.3%) in 2020. The main reasons for non-enrollment identified were refusal of treatment due to financial difficulties, loss to follow-up or death before starting treatment. Multivariate analysis showed that the elderly patients aged 65 or above (OR = 2.7, CI: 1.997-3.614), new patients (OR = 0.7, CI: 0.607-0.867), conventional DST used for confirmatory diagnosis of RR-TB (OR = 1.9, CI: 1.620-2.344) and diagnosis of RR-TB being conducted by the RR-TB care facilities at the prefecture and municipal level (OR = 4.4, CI: 3.608-5.250) have implications on RR-TB non-enrollment. As a comprehensive RR-TB model was implemented in Yunnan with scaled up use of molecular test for rapid detection of RR-TB, initial screening of RR-TB were decentralized to the county- and district-level to strengthen rapid, early detection of RR-TB, achieving a higher coverage of screening in the end. However, there remains a major gap in enrollment of RR-TB. The main barriers include: limited knowledge and awareness of RR-TB and financial burdens among patients, delayed diagnosis, loss to follow-up, difficulties in self care and travel for elderly patients, and limited capacity of clinical management at the lower-level RR-TB care facilities. The situation of the RR-TB epidemic in Yunnan could be improved and contained as soon as possible by continuous strengthening of the comprehensive, patient-centered model with targeted interventions coordinated through multi-sectoral engagement to improve enrollment of RR-TB patients.