Association between Mycobacterium tuberculosis genotype and diabetes mellitus/hypertension: a molecular study.

BACKGROUND: A paucity of studies focused on the genetic association that tuberculosis (TB) patients with non-communicable diseases (NCDs) are more likely to be infected with Mycobacterium tuberculosis (MTB) with more potent virulence on anti-TB drug resistance than those without NCDs. The study aimed to document the predominant genotype, determine the association between MTB genotypes and NCD status and drug resistance. METHODS: We conducted a molecular study in 105 TB patients based on a cross-sectional study focused on the comorbid relationship between chronic conditions and TB among 1773 subjects from September 1, 2019 to August 30, 2020 in Guizhou, China. The participants were investigated through face-to-face interviews, followed by NCDs screening. The DNA of MTB isolates was extracted prior to genotyping using 24 loci MIRU-VNTR. The subsequent evaluations were performed by phylogenetic trees, combined with tests of statistical power, Chi-square or Fisher and multivariate logistic regression analysis. RESULTS: The Beijing family of Lineage 2 (East Asia) was the predominant genotype accounting for 43.8% (46/105), followed by Lineage 4 (Euro-America) strains, including Uganda I (34.3%, 36/105), and the NEW-1 (9.5%, 10/105). The proportion of Beijing strain in patients with and without NCDS was 28.6% (8/28) and 49.4% (38/77), respectively, with a statistical power test value of 24.3%. No significant association was detected between MTB genotype and NCD status. A low clustering rate (2.9%) was identified, consisting of two clusters. The rates of global, mono-, poly- and multi-drug resistance were 16.2% (17/105), 14.3% (15/105), 1.0% (1/105) and 4.8% (5/105), respectively. The drug-resistant rates of rifampicin, isoniazid, and streptomycin, were 6.7% (7/105), 11.4% (12/105) and 5.7% (6/105), respectively. Isoniazid resistance was significantly associated with the Beijing genotype of Lineage 2 (19.6% versus 5.1%). CONCLUSIONS: The Lineage 2 East Asia/Beijing genotype is the dominant genotype of the local MTB with endogenous infection preponderating. Not enough evidence is detected to support the association between the MTB genotype and diabetes/hypertension. Isoniazid resistance is associated with the Lineage 2 East Asia/Beijing strain.

Risk factors of multidrug-resistant tuberculosis in China: A meta-analysis.

BACKGROUND: Multidrug-resistant tuberculosis (MDR-TB) brings major challenges to the health care workers (HCWs). This study is to determine the risk factors for MDR-TB, latent tuberculosis infection (LTBI), and tuberculosis (TB) disease among HCWs in China. METHODS: A meta-analysis was conducted to evaluate the risk factors for MDR-TB, LTBI, and TB disease among HCWs using a random-effects model, and the pooled odds ratios (ORs) with 95% confidence interval (CI) were used as effect indicators. RESULTS: We identified 46 eligible studies and found eight factors were associated with MDR. The ORs with 95% CI are migrant population 1.96 (95% CI, 1.50-2.57), low family income 2.23 (95% CI, 1.74-2.85), retreatment 7.22 (95% CI, 5.63-9.26), anti-TB treatment history 5.65 (95% CI, 4.80-6.65), multiple episodes of treatment 3.28 (95% CI, 2.60-4.13), adverse reactions 3.48 (95% CI, 2.54-4.76), interrupted treatment 3.18 (95% CI, 2.60-3.89), and lung cavities 1.42 (95% CI, 1.14-1.77). Work duration as a HCW for 5 years and above increased the risk of LTBI and TB. HCWs aged 30 years and above were more susceptible to TB (OR = 1.70, 95% CI: 1.37-2.09). CONCLUSION: The risk factors for MDR-TB in China are possibly migrant population, low family income, retreatment, anti-TB treatment history, adverse reactions, interrupted treatment, and lung cavities. Longer work duration and greater age are risk factors for LTBI and TB among HCWs.

Lot-to-lot consistency, immunogenicity and safety of a quadrivalent split virion inactivated influenza vaccine in healthy population aged 9-59 years: A randomized, double-blind, controlled, phase IV clinical trial.

OBJECTIVES: This study was to assess the lot-to-lot consistency, immunogenicity and safety of three manufacturing lots of a quadrivalent inactivated influenza vaccine (IIV4). METHODS: A randomized, double-blind, phase IV clinical trial was conducted in healthy children, adolescents and adults aged 9-59 years in Guizhou Province, China. Eligible participants were enrolled and randomized into three groups in a ratio of 1:1:1 to receive a single dose of one of three manufacturing lots of IIV4. Serum samples were collected before and 28 days after vaccination for hemagglutination inhibition (HI) antibody testing. Safety data were collected for up to 28 days after vaccination. The primary objective was to evaluate the lot-to-lot consistency of immune response as assessed by the geometric mean titer (GMT) of HI antibody at 28 days after vaccination. RESULTS: Between November 27, 2022 and December 18, 2022, 1260 eligible participants were enrolled, with similar participant demographics among groups. Immune responses after vaccination were comparable across groups, with the 95% confidence intervals (CIs) of GMT ratios for all 4 strains falling into the equivalence criterion of (0.67, 1.5). The seroconversion rates (SCRs) and seroprotection rates (SPRs) met the US Center or Biologics Evaluation and Research (CBER) criteria for all strains for each lot (lower limit of 95% CI of SCR ≥ 40% and SPR ≥ 70%). The incidences of solicited and unsolicited adverse reactions were similar among three groups, most of which (91.9%) were mild or moderate in severity. A total of 11 serious adverse events were reported during the study, and all were considered unrelated to vaccination. CONCLUSION: The three manufacturing lots of IIV4 demonstrated consistent immunogenicity. IIV4 can elicit satisfactory immune responses for all four strains and no safety concerns were identified. CLINICAL TRIAL REGISTRATION: Identifier No. NCT05512494.

Immunogenicity and safety of quadrivalent inactivated influenza vaccine in children aged 6 to 35 months: A systematic review and meta-analysis.

Evidence of the immunogenicity and safety of quadrivalent inactivated influenza vaccine in children aged 6 to 35 months has been emerging. To evaluate the immunogenicity and safety of quadrivalent inactivated influenza vaccine in children aged 6 to 35 months in a systematic review and meta-analysis. This meta-analysis included 12 studies with 6722 participants receiving QIV, 3575 participants receiving TIV, 4249 participants receiving full-dose QIV (F-QIV), and 3722 participants receiving half-dose QIV (H-QIV). Among children aged 6 to 35 months, QIV produces a better Immunogenicity against influenza B vaccine strains not contained in TIV. However, injection site reaction was more common for QIV, F-QIV showed superior efficacy for the B lineage, but fever and injection site pain was more frequently reported for F-QIV than H-QIV. These data support the immunogenicity and safety of quadrivalent inactivated influenza vaccine among children aged 6 to 35 months.

Development of CRISPR/Cas12b-Based Multiple Cross Displacement Amplification Technique for the Detection of Mycobacterium tuberculosis Complex in Clinical Settings.

Tuberculosis (TB) is a chronic infectious disease with high mortality caused by the Mycobacterium tuberculosis complex (MTC). Its clinical symptoms include a prolonged cough with mucus, pleuritic chest pain, hemoptysis, etc., and predominant complications such as tuberculous meningitis and pleural effusion. Thus, developing rapid, ultrasensitive, and highly specific detection techniques plays an important role in controlling TB. Here, we devised CRISPR/CRISPR-associated 12b nuclease (CRISPR/Cas12b)-based multiple cross displacement amplification technique (CRISPR-MCDA) targeting the IS6110 sequence and used it to detect MTC pathogens. A newly engineered protospacer adjacent motif (PAM) site (TTTC) was modified in the linker region of the CP1 primer. In the CRISPR-MCDA system, the exponentially amplified MCDA amplicons with the PAM sites can guide the Cas12b/gRNA complex to quickly and accurately recognize its target regions, which successfully activates the CRISPR/Cas12b effector and enables ultrafast trans-cleavage of single-stranded DNA reporter molecules. The limit of detection of the CRISPR-MCDA assay was 5 fg/µL of genomic DNA extracted from the MTB reference strain H37Rv. The CRISPR-MCDA assay successfully detected all examined MTC strains and there was no cross-reaction with non-MTC pathogens, confirming that its specificity is 100%. The entire detection process can be completed within 70 min using real-time fluorescence analysis. Moreover, visualization detection (under UV light) was also designed to verify the results, eliminating the use of specialized instruments. In conclusion, the CRISPR-MCDA assay established in this report can be used as a valuable detection technique for MTC infection. IMPORTANCE The Mycobacterium tuberculosis complex pathogen is a crucial infectious agent of tuberculosis. Hence, improving the capability of MTC detection is one of the most urgently required strategies for preventing and controlling TB. In this report, we successfully developed and implemented CRISPR/Cas12b-based multiple cross displacement amplification targeting the IS6110 sequence to detect MTC pathogens. These results demonstrated that the CRISPR-MCDA assay developed in this study was a rapid, ultrasensitive, highly specific, and readily available method which can be used as a valuable diagnostic tool for MTC infection in clinical settings.

Ultrafast, One-Step, Label-Based Biosensor Diagnosis Platform for the Detection of Mycobacterium tuberculosis in Clinical Applications.

Tuberculosis (TB) is a chronic infectious disease caused by the etiological agent Mycobacterium tuberculosis (MTB). Because the majority of TB patients come from poor economic backgrounds, the development of a simple, specific, low-cost, and highly sensitive detection method for the pathogen is extremely important for the prevention and treatment of this disease. In the current study, an efficient detection method for visual, rapid, and highly sensitive detection of MTB utilizing multiplex loop-mediated isothermal amplification combined with a label-based lateral flow immunoassay biosensor (mLAMP-LFIA) was developed. Three specific primer sets targeting the MTB genes IS6110 and mpb64 were successfully designed and synthesized for the LAMP assay. The optimal reaction conditions for the mLAMP-LFIA assay were confirmed to be 67 °C for 40 min. The mLAMP amplicons were intuitively verified using the LFIA biosensor within 5 min. The entire process, including clinical sample processing, amplification reaction, and product verification, was completed within 80 min. The limit of detection of the mLAMP-LFIA assay established in the current study was 100 fg per reaction for the genomic DNA of MTB H37Rv. The analytical specificity of the mLAMP-LFIA assay was one hundred percent, and no cross-reactions with non-target strains were detected. Compared with the GeneXpert test, the sensitivity of mLAMP-LFIA for 148 clinical specimens was 100% (97/97), and the specificity was 98.04% (50/51) in the preliminary evaluation of the clinical application. Hence, the mLAMP-LFIA method, targeting the IS6110 and mpb64 genes, is an ultrafast, one-step, low-cost, and highly sensitive detection method that could be used as a screening and/or diagnostic tool for MTB in the clinical setting, basic science laboratories, and especially in resource-poor regions.

Immune-Boosting Effect of the COVID-19 Vaccine: Real-World Bidirectional Cohort Study.

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.

Comparison on Major Gene Mutations Related to Rifampicin and Isoniazid Resistance between Beijing and Non-Beijing Strains of Mycobacterium tuberculosis: A Systematic Review and Bayesian Meta-Analysis.

Objective: The Beijing strain of Mycobacterium tuberculosis (MTB) is controversially presented as the predominant genotype and is more drug resistant to rifampicin and isoniazid compared to the non-Beijing strain. We aimed to compare the major gene mutations related to rifampicin and isoniazid drug resistance between Beijing and non-Beijing genotypes, and to extract the best evidence using the evidence-based methods for improving the service of TB control programs based on genetics of MTB. Method: Literature was searched in Google Scholar, PubMed and CNKI Database. Data analysis was conducted in R software. The conventional and Bayesian random-effects models were employed for meta-analysis, combining the examinations of publication bias and sensitivity. Results: Of the 8785 strains in the pooled studies, 5225 were identified as Beijing strains and 3560 as non-Beijing strains. The maximum and minimum strain sizes were 876 and 55, respectively. The mutations prevalence of rpoB, katG, inhA and oxyR-ahpC in Beijing strains was 52.40% (2738/5225), 57.88% (2781/4805), 12.75% (454/3562) and 6.26% (108/1724), respectively, and that in non-Beijing strains was 26.12% (930/3560), 28.65% (834/2911), 10.67% (157/1472) and 7.21% (33/458), separately. The pooled posterior value of OR for the mutations of rpoB was 2.72 ((95% confidence interval (CI): 1.90, 3.94) times higher in Beijing than in non-Beijing strains. That value for katG was 3.22 (95% CI: 2.12, 4.90) times. The estimate for inhA was 1.41 (95% CI: 0.97, 2.08) times higher in the non-Beijing than in Beijing strains. That for oxyR-ahpC was 1.46 (95% CI: 0.87, 2.48) times. The principal patterns of the variants for the mutations of the four genes were rpoB S531L, katG S315T, inhA-15C > T and oxyR-ahpC intergenic region. Conclusion: The mutations in rpoB and katG genes in Beijing are significantly more common than that in non-Beijing strains of MTB. We do not have sufficient evidence to support that the prevalence of mutations of inhA and oxyR-ahpC is higher in non-Beijing than in Beijing strains, which provides a reference basis for clinical medication selection.

Gradient association between pulmonary tuberculosis and diabetes mellitus among households with a tuberculosis case: a contact tracing-based study.

Pulmonary tuberculosis (PTB) and diabetes mellitus (DM) remain high morbidity and mortality, especially when they are comorbid with each other. Screening for diabetes mellitus in tuberculosis is essential as the incidence and mortality of DM in the population with PTB are higher than in the general people. We aimed to examine the gradient association of tuberculosis on developing DM, the additional yield and the number needed to screen (NNS) to find a new diabetes case. A cross-sectional study was conducted on 801 tuberculosis cases and 972 household contacts in Guizhou, China, from April 2019 to October 2020. After screening for PTB among contacts, all participants were screened for DM and interviewed. Kendall's tau-b test and proportional odds logistic regression analysis were applied to identify the gradient associations. Among the 1773 subjects, the additional yield of screening was 21.8%. The NNSs of the non-PTB group, the sputum-culture negative and positive groups were 50, 60 and 113, respectively. The gradient incremental establishment of DM and PTB were positively correlated. The general trend on the gradient of DM significantly increased with the gradient increase of PTB. Age 35 years and over, excessive edible oil intake and DM family history were identified as significant predictors of diabetes. Integrated screening for DM targeted to different gradients of PTB combined with associated factors is necessitated to achieve a higher additional yield.

Yield, NNS and prevalence of screening for DM and hypertension among pulmonary tuberculosis index cases and contacts through single time screening: A contact tracing-based study.

INTRODUCTION: Diabetes mellitus (DM), hypertension and pulmonary tuberculosis (PTB) are catastrophic illnesses that collectively lead to increased mortality and premature death. However, the size of the problem and the appropriate approach to deal with the burden is still unclear. We aimed to evaluate the yield, number needed to screen (NNS) to prevent one death or adverse event for screening DM and hypertension and assess the prevalence and contributors to DM and/or hypertension. METHODS: Based on PTB contact tracing, a cross-sectional study was conducted among 801 PTB index cases and 972 household contacts from April 2019 to October 2020 in Guizhou, China. All the participants were screened for DM and hypertension. The yield was calculated as the proportion of newly detected cases among the study subjects, excluding known cases. The NNS was computed by dividing the number needed to treat for risk factors by the prevalence of the unrecognized diseases. The univariate and multivariate logistic regression analyses were applied to determine the independent predictors of DM and/or hypertension. RESULTS: Of the 1,773 participants, the prevalence of DM and hypertension was 8.7% (70/801) and 15.2% (122/801) in the PTB patients, 3.2% (31/972) and 14.0% (136/972) in the contacts, respectively. The prevalence of DM and/or hypertension was 21.2% (170/801) among the PTB patients and 15.4% (150/972) among their contacts. The screening yields to detect new cases of DM and hypertension among PTB patients were 1.9% and 5.2%, and that in the contacts were 0.8% and 4.8%, respectively. The NNS for DM was 359 for the PTB cases and 977 for the contacts, 299 for PTB cases and 325 for hypertension, respectively. Older age, under or overweight and obesity, family history hypertension and earlier diagnosis of other chronic conditions were the independent predictors for DM and/or hypertension among both PTB cases and their contacts. CONCLUSION: Screening for DM and hypertension should be mandated in PTB patients and their household contacts to disclose undetected cases of these two conditions during TB contact tracing, which might reduce the potential cardiovascular disease deaths.

Rapid and Visual Differentiation of Mycobacterium tuberculosis From the Mycobacterium tuberculosis Complex Using Multiplex Loop-Mediated Isothermal Amplification Coupled With a Nanoparticle-Based Lateral Flow Biosensor.

Tuberculosis (TB) is a chronic infectious disease mainly caused by Mycobacterium tuberculosis (MTB), but other members of the Mycobacterium tuberculosis complex (MTBC), especially Mycobacterium bovis (pyrazinamide-resistant organisms), may also be involved. Thus, the ability to rapidly detect and identify MTB from other MTBC members (e.g., M. bovis, Mycobacterium microti, Mycobacterium africanum) is essential for the prevention and treatment of TB. A novel diagnostic method for the rapid detection and differentiation of MTB, which employs multiplex loop-mediated isothermal amplification (mLAMP) combined with a nanoparticle-based lateral flow biosensor (LFB), was established (mLAMP-LFB). Two sets of specific primers that target the IS6110 and mtp40 genes were designed according to the principle of LAMP. Various pathogens were used to optimize and evaluate the mLAMP-LFB assay. The optimal conditions for mLAMP-LFB were determined to be 66°C and 40 min, and the amplicons were directly verified by observing the test lines on the biosensor. The LAMP assay limit of detection (LoD) was 125 fg per vessel for the pure genomic DNA of MTB and 4.8 × 103 CFU/ml for the sputum samples, and the analytical specificity was 100%. In addition, the whole process, including the clinical specimen processing (35 min), isothermal amplification (40 min), and result confirmation (1-2 min), could be completed in approximately 80 min. Thus, mLAMP-LFB is a rapid, reliable, and sensitive method that is able to detect representative members of MTBC and simultaneously differentiate MTB from other MTBC members, and it can be used as a potential screening tool for TB in clinical, field, and basic laboratory settings.

Immunogenicity and safety of a live attenuated varicella vaccine in children 1-12 years of age: A randomized, blinded, controlled, non-inferiority phase 3 clinical trial.

OBJECTIVES: To evaluate the immunogenicity and safety of a live attenuated varicella vaccine produced using a cell factory process. METHODS: In this randomized, blinded, controlled, non-inferiority phase 3 clinical trial conducted in Guizhou, healthy children aged 1-12 years were randomly assigned in a 2: 1 ratio to receive one dose of experimental or control vaccine. Physical examination and first blood collection were performed preimmunization on day 0. Diary cards were collected after day 15. Contact cards and second blood samples were collected on day 30. The primary immunogenicity endpoint was the positive conversion rate of the anti-varicella virus antibody at 30 days postimmunization in susceptible children. Secondary endpoints were the fourfold increase rate, positive conversion rate, geometric mean titer, and geometric mean increase at 30 days after immunization in the total cohort. RESULTS: Of the 900 children assessed for eligibility, 894 received an experimental or control vaccine. Both the full analysis and safety analysis sets included 894 subjects. The seroconversion rate in the susceptible population was 95.84% in the experimental and 94.76% in the control group. The lower limit of the 95% confidence interval difference was -2.37%, which was greater than the non-inferiority margin set by the program (-10%). No significant difference in solicited adverse reactions was found between the groups. Within 6 months postimmunization, a total of 24 serious adverse events were reported, none related to the studied vaccine. CONCLUSION: The live attenuated varicella vaccine produced using a cell factory process was highly immunogenic, safe, and non-inferior to the product in the market. Further studies need to be implemented in the immune persistence, the epidemiological effectiveness and the rare adverse reactions.

Investigation and analysis of Galerina sulciceps poisoning in a canteen.

INTRODUCTION: Guizhou Province in China has an abundant resource of wild mushrooms, including numerous poisonous species which contain various toxins. The mortality rate from wild mushroom poisoning has been high in this area in recent years. Galerina sulciceps is a dangerously toxic mushroom which can be fatal if ingested. METHODS: we report on an epidemiological investigation of G. sulciceps poisoning which occurred in Duyun City of Guizhou Province. The characteristics of this species, its toxicity, observed clinical features, laboratory data, treatment modality, and prognosis were investigated in order to provide a reference point for the prevention and treatment of this kind of mushroom poisoning. RESULTS: Thirteen employees showed toxic symptoms after ingesting wild mushrooms the previous day in a company canteen. Clinical manifestation varied from gastroenteritis to hepatic and renal dysfunction. Most of the 13 patients presented with nausea, vomiting, abdominal pain, diarrhea, and elevated levels of biochemical indices of hepatic and renal function, during which transaminase concentration peaked within 48-72 h. At 48 hours post-ingestion, all patients had hemodialysis, in addition to supportive care for hepatic and renal injury with oral Silibinin and Shenshuaining. All acute renal injury had resolved by day 10, and liver transaminases had trended toward normal in all patients and they were discharged. At follow-up in 30 days, both liver and renal function had completely recovered in all. CONCLUSION: This poisoning occurs as a result of unintentional consumption of G. sulciceps, which is relatively rare in mushroom poisonings. All patients recovered fully after timely diagnosis and treatment. To prevent wild mushroom poisoning, the best preventive measure is to educate the public not to gather and eat any unidentified wild mushrooms.