Guanghong Yang

Mycobacterium tuberculosis complex (MTBC), the primary causative agent of pulmonary tuberculosis (PTB), shares pathological features with nontuberculous mycobacteria (NTM). Some NTM species act as opportunistic pathogens that can cause NTM lung disease, often leading to diagnostic confusion with PTB. Given the persistent global burden of tuberculosis and the rising incidence of NTM infections worldwide, there is an urgent need for rapid, highly sensitive diagnostic techniques capable of distinguishing MTBC and NTM at initial clinical presentation. In this study, we developed a novel diagnostic platform that integrates loop-mediated isothermal amplification (LAMP) with nanoparticle-based lateral flow biosensors (LFB), and real-time (RT) fluorescence analysis for simultaneous detection of MTBC and NTM. Target-specific primers were designed for IS6110 (MTBC-specific) and 16S rDNA (pan-mycobacterial), and the corresponding probes were multi-labeled: FAM-biotin/digoxigenin-biotin for LFB capture, and FAM-BHQ1/Cy5-BHQ2 for real-time fluorescence monitoring. After systematic optimization, the LFB–RT–LAMP assay performed best at 67°C for 50 min. Analytical evaluation using genomic DNA from MTB reference strain H37Rv, 22 mycobacterial strains, 13 clinically relevant non-mycobacterial pathogens, and 80 respiratory specimens showed the following: (i) a limit of detection (LoD) of 576 fg/reaction for purified MTB DNA; (ii) 100% detection rate for the tested mycobacteria; (iii) 100% specificity against non-target pathogens; (iv) 100% clinical concordance with composite diagnostic PCR-based methods; and (v) a turnaround time of IS6110 ) and NTM screening (via 16S rDNA) in a single assay. The method achieves a turnaround time of under 60 min, a sensitivity of 576 fg/reaction, and 100% specificity against non-mycobacterial pathogens, demonstrating potential for point-of-care use in clinical and field settings. IMPORTANCE: Tuberculosis (TB) and nontuberculous mycobacteria (NTM) lung disease present with similar symptoms but require completely different treatment regimens. Rapid and accurate differentiation between these pathogens at initial presentation remains a critical unmet need, as current molecular tests primarily focus on TB and are not designed for NTM screening. In this study, we developed a simple, rapid, and equipment‑free diagnostic platform that distinguishes TB from NTM within 60 min using a lateral flow biosensor, alongside a real‑time fluorescence option for laboratory use. The assay simultaneously targets IS6110 ( Mycobacterium tuberculosis complex [MTBC]‑specific) and 16S rDNA (pan‑mycobacterial), enabling species‑level identification of MTBC and genus‑level screening of NTM in a single reaction, achieving 100% accuracy on clinical samples with a limit of detection of 576 fg (~120 genome copies) per reaction. This dual‑modal approach offers practical solutions for both well‑equipped hospitals and remote clinics, addressing a critical gap in TB and NTM diagnostics.

Background: As the SARS-CoV-2 attenuates and antibodies from the COVID-19 vaccine decline, long-term attention should be paid to the durability of primary booster administration and the preventive effect of the second or multiple booster doses of the COVID-19 vaccine. Objective: This study aimed to explore the durability of primary booster administration and the preventive effect of second or multiple booster doses of the COVID-19 vaccine. Methods: We established a bidirectional cohort in Guizhou Province, China. Eligible participants who had received the primary booster dose were enrolled for blood sample collection and administration of the second booster dose. A retrospective cohort for the time of administration was constructed to evaluate antibody attenuation 6-12 months after the primary booster dose, while a prospective cohort on the vaccine effect of the second booster dose was constructed for 4 months after the second administration. Results: Between September 21, 2022, and January 30, 2023, a total of 327 participants were included in the final statistical analysis plan. The retrospective cohort revealed that approximately 6-12 months after receiving the primary booster, immunoglobulin G (IgG) slowly declined with time, while immunoglobulin A (IgA) remained almost constant. The prospective cohort showed that 28 days after receiving the second booster, the antibody levels were significantly improved. Higher levels of IgG and IgA were associated with better protection against COVID-19 infection for vaccine recipients. Regarding the protection of antibody levels against post–COVID-19 symptoms, the increase of the IgG had a protective effect on brain fog and sleep quality, while IgA had a protective effect on shortness of breath, brain fog, impaired coordination, and physical pain. Conclusions: The IgG and IgA produced by the second booster dose of COVID-19 vaccines can protect against SARS-CoV-2 infection and may alleviate some post–COVID-19 symptoms. Further data and studies on secondary booster administration are required to confirm these conclusions.

Background: With the rapid spread of the mpox epidemic, cases have emerged in multiple countries, mainly among men who have sex with men. Because of the connectedness of today’s world, countries have to be prepared to face risks in advance. Therefore, this study aimed to investigate awareness of mpox-related knowledge among men who have sex with men in China. Methods: With the assistance of the social organizations of men who have sex with men, a cross-sectional survey of men who have sex with men in China was conducted through an online questionnaire between July 1 and July 18, 2022. A nationwide sample of Chinese men who have sex with men (N = 3,257) was recruited. Results: Only 36.9% of participants had mpox-related knowledge. Awareness of mpox-related knowledge among respondents was positively associated with those in older age groups (33 to 42 years and 51 years or older) (adjusted odds ratio [AOR] = 1.31; 95% confidence interval [CI]: 1.03–1.67, AOR = 1.61; 95% CI: 1.16–2.24; respectively), married (AOR = 1.55; 95% CI: 1.09–2.19), and those with a graduate degree or above (AOR = 2.14; 95% CI: 1.11–4.13), while negatively associated with those living in the western parts of China (AOR = 0.74; 95% CI: 0.60–0.92), and those who were unsure of their history of Human Immunodeficiency Virus (HIV) status (AOR = 0.44; 95% CI: 0.30–0.63). Conclusion: Mpox-related knowledge is fairly low among men who have sex with men in China. China needs to spread knowledge to the public through multiple channels, especially in key populations (men who have sex with men, HIV-infected, etc.), and take preventive measures to effectively avoid outbreaks of mpox.