OBJECTIVE To explore the diagnostic value of combined detection of droplet digital polymerase chain reaction(ddPCR),C-reactive protein(CRP),procalcitonin(PCT)and neutrophil-to-lymphocyte ratio(NLR)for bacterial bloodstream infections.METHODS Patients with suspected bloodstream infections admitted to Jiangbin Hospital of Guangxi Zhuang Autonomous Region from Jan.2023 to Jun.2024 were selected as the study subjects,with a total of 993 specimens from 543 patients included.Based on ddPCR and blood culture(BC)results,the pa-tients were divided into BC+and/or ddPCR+group(424 specimens)and ddPCR-/BC-group(569 specimens).The bacterial species detected by ddPCR were further classified into single infection group(258 speci-mens),mixed infection group(160 specimens)and ddPCR-group(575 specimens).The levels of CRP,PCT and NLR were compared among the groups.Receiver operating characteristic(ROC)curves were used to evaluate the diagnostic efficacy of ddPCR,CRP,PCT and NLR separately and in combination for bacterial bloodstream in-fections.RESULTS The levels of CRP,PCT and NLR in the BC+and/or ddPCR+group were 71.61(37.00,108.81)mg/L,1.74(0.47,7.93)ng/ml and 9.82(5.53,18.07),respectively,which were higher than those in the ddPCR-/BC-group(P＜0.001).I n the ddPCR mixed infection group,the levels of CRP and PCT were 88.02(42.90,112.39)mg/L and 2.83(0.89,12.35)ng/ml,respectively,which were higher than those in the single infection group(P＜0.05).The qualitative results of ddPCR were better at predicting bloodstream infec-tions(OR=15.279,95％CI:6.525～35.776,P＜0.001).According to ROC curve analysis,ddPCR had the lar-gest area under the curve(AUC)among the single detection indicator(0.759),followed by PCT(0.732).The AUC for the combination of ddPCR qualitative results with CRP,PCT and NLR was 0.830,indicating that the di-agnostic performance of the combined detection was better than that of any single indicator.CONCLUSIONS The combination of ddPCR with PCT,CRP and NLR can improve the accuracy of diagnosing bacterial bloodstream in-fections.Compared with BC,ddPCR has higher sensitivity and can quickly identify the types and concentrations of pathogens in bloodstream infections.

OBJECTIVE To explore the diagnostic value of combined detection of droplet digital polymerase chain reaction(ddPCR),C-reactive protein(CRP),procalcitonin(PCT)and neutrophil-to-lymphocyte ratio(NLR)for bacterial bloodstream infections.METHODS Patients with suspected bloodstream infections admitted to Jiangbin Hospital of Guangxi Zhuang Autonomous Region from Jan.2023 to Jun.2024 were selected as the study subjects,with a total of 993 specimens from 543 patients included.Based on ddPCR and blood culture(BC)results,the pa-tients were divided into BC+and/or ddPCR+group(424 specimens)and ddPCR-/BC-group(569 specimens).The bacterial species detected by ddPCR were further classified into single infection group(258 speci-mens),mixed infection group(160 specimens)and ddPCR-group(575 specimens).The levels of CRP,PCT and NLR were compared among the groups.Receiver operating characteristic(ROC)curves were used to evaluate the diagnostic efficacy of ddPCR,CRP,PCT and NLR separately and in combination for bacterial bloodstream in-fections.RESULTS The levels of CRP,PCT and NLR in the BC+and/or ddPCR+group were 71.61(37.00,108.81)mg/L,1.74(0.47,7.93)ng/ml and 9.82(5.53,18.07),respectively,which were higher than those in the ddPCR-/BC-group(P＜0.001).I n the ddPCR mixed infection group,the levels of CRP and PCT were 88.02(42.90,112.39)mg/L and 2.83(0.89,12.35)ng/ml,respectively,which were higher than those in the single infection group(P＜0.05).The qualitative results of ddPCR were better at predicting bloodstream infec-tions(OR=15.279,95％CI:6.525～35.776,P＜0.001).According to ROC curve analysis,ddPCR had the lar-gest area under the curve(AUC)among the single detection indicator(0.759),followed by PCT(0.732).The AUC for the combination of ddPCR qualitative results with CRP,PCT and NLR was 0.830,indicating that the di-agnostic performance of the combined detection was better than that of any single indicator.CONCLUSIONS The combination of ddPCR with PCT,CRP and NLR can improve the accuracy of diagnosing bacterial bloodstream in-fections.Compared with BC,ddPCR has higher sensitivity and can quickly identify the types and concentrations of pathogens in bloodstream infections.

Objective To investigate the mutation characteristics and influencing factors of ethambutol(EMB)resistance gene of Mycobacterium tuberculosis in Guangxi Zhuang Autonomous region,and to provide evidence for molecular diagnosis and clinical treatment of tuberculosis.Methods A total of 655 strains of Myco-bacterium tuberculosis(52 ethambutol resistant strains and 603 ethambutol sensitive strains)were collected continuously from 30 TB drug resistance monitoring sites in Guangxi in 2018-2019,and the mutation characteristics and influencing factors of ethambutol resistant genes were analyzed by whole genome sequencing.Results Among 655 strains of Mycobacterium tuberculosis,54 strains had ethambutol drug resistance gene mutation,the mutation rate was 8.24%(54/655).Among 52 EMB-resistant strains detected by proportional method,21 had gene mutation,the mutation rate was 40.38%(21/52),and 33 of 603 EMB-sensitive strains had gene mutation,the mutation rate was 5.47%(33/603).The gene mutation rate in drug-resistant strains was higher than that in sensitive strains(χ2 = 77.133,P = 0.000).The coincidence rate of EMB drug resistance phenotype and gene mutation was 40.38%(21/52),and the results of the two tests were not highly consistent(Kappa = 0.343,P<0.001).The mutant genes of 54 strains were embA,embB and embC,and there were 20 mutant forms,among which 29 were mutated at unit point,accounting for 53.70%(29/54),and 25 were mutated at joint site,accounting for 46.30%(25/54).Among the unit point mutations,embB306(35.19%)had the highest mutation proportion,followed by embB497(5.56%)and embB406(3.70%).Among the joint site mutations,embC270+embB378 had the highest mutation proportion(22.22%),The second was embB306+embA-12(3.70%).Gender,anti-tuberculosis treatment history,genotype and MDR might be related to EMB gene mutation(χ2 = 9.388,P = 0.004;χ2 = 27.084,P = 0.000;χ2 = 6.671,P = 0.010;χ2 = 68.826,P = 0.000).Multivariate logistic regression analysis showed that male(OR = 6.150),retreatment(OR = 2.636)and multidrug resistance(OR = 7.333)may be risk factors for EMB resistance gene mutation,and Beijing genotype may be a protective factor for EMB resistance gene mutation(OR = 0.511).Conclusion EMB resistance of Mycobacterium tuberculosis is related to embA,embB and embC gene mutations,and the incidence of EMB resistance phenotype is not high.For male,retreatable,MDR-resistant,and non-Beijing genotype TB patients,attention should be paid to the mutation of the EMB resistance gene.

Objective To investigate the genetic diversity of variable number tandem repeats (VNTR) of Mycobacterium tuberculosis (M,ib) isolates and their impact factors on transmission.Methods A total of 1 310 patients with smear-positive pulmonary tuberculosis registered during 2013 from 30 counties of drug-resistance monitoring sites in Guangxi were enrolled.The VNTR genotyping technique was used to identify and analyze M.tb genotypes.Count data was analyzed by Chi-square test using descriptive statistical method,unconditioned Logistic regression was used to analyze the characteristics of gene clusters.Results A total of 1 310 strains of M.tb were classified into 964 genotypes,of which 779 strains were classified into a single gene cluster,531 strains were classified into 185 clusters.Each cluster contained 2 to 40 strains (mean 6.6).Patients aged between 41 and 60 years old had a highest gene cluster proportion (45.5％).Seven hundred and forty-six strains were Beijing genotype,while 564 were nonBeijing genotype,among which 7.4％ was original Beijing genotype.The estimated recent infection (cluster rate) was 26.41％.The cluster rate was higher in sensitive strains (24.40％) than in drugresistant strains (9.55 ％,x2 =23.621,P =0.000) and multi-drug resistant (MDR) strains (8.97 ％,x2=-9.675,P=0.002),and that was higher in Beijing genotype (28.69％) than Non-Beijing genotype (23.40％,x2 =4.610,P=0.032).The clusters proportion in sensitive strain (37.43％) was higher than drug-resistant strains (14.09％) and MDR strains (15.38％),and that was higher in Beijing genotype (40.88％) than Non-Beijing genotype (37.77％).Conclusions M.tb genetic polymorphisms display highly diversity in Guangxi region.Sensitive strains and Beijing genotype strains are the main epidemic strains.