Objective To establish a rapid immunological detection method for MPT64 protein based on red microspheres and select highly-sensitive and highly-specific antibody pairs.Methods A His-tagged prokaryotic expression vector was constructed for expression of MPT64 protein that was used to immunize New Zealand white rabbits after purification and validation.Peripheral blood mononuclear cells(PBMCs)were isolated from the rabbits,and antigen-specific B cells expressing antibodies were sorted using single B-cell flow cytometry.mRNA in B cells was reverse-transcribed into cDNA,and paired antibody heavy-and light-chain sequences were amplified via nested PCR.Expression vectors were constructed,and recombinant antibodies were produced in Expi293F cells.Fluorescent immunochromatography was employed to screen for matched antibody pairs.The selected antibodies were used to establish a rapid detection method based on red microsphere immunochromatography.Results Ten high-affinity monoclonal antibodies against MPT64 were generated.Two antibody pairs were selected for MPT64 immunodetection that reached a sensitivity of 0.0125 ng/mL.Conclusion High-affinity rabbit monoclonal antibodies against MPT64 are obtained via single B-cell technology,and a rapid red microspheres-based immunodetection method is established,enabling highly sensitive detection of Mycobacterium tuberculosis MPT64 protein.

In recent years,sudden public health events caused by group A streptococcus infections have been emerging,imposing a huge economic burden on society.The development of group A streptococcus vaccines has been an area of great interest to scientists.There is currently a wide range of vaccines in different stages of development.However,no mature and usable vaccines are available so far.This paper reviewed the current research on group A streptococcus vaccines in general and on M-protein recombinant protein vaccines,non-M-protein recombinant protein vaccines,and polysaccharide conjugate vaccines in particular.

Aim To investigate the gender differences in electrophysiology and neuroanatomy of myelinated and unmyelinated visceral and baroreceptor afferent neurons(VANs and ABNs) of adult rats.Methods VANs and ABNs were isolated enzymatically and Vagus-nodose slice preparation was also applied in this study.For identification of ABNs,aortic depressor nerve(ADN) was labeled using fluorescent dye.Whole-cell patch technique was used to record action potential(AP).Electronic microscopy was selected for morphological analysis of ADN.Results(1) A-and C-type VANs were identified and significant differences of AP discharge profiles between female and male were not established;(2) except for the traditionally classified A-and C-types,myelinated Ah-type VGNs were also identified with faster conduction velocity,lower firing threshold,and higher neuronal excitability.Importantly,these Ah-types were found in female rats with a similar frequency like A-types but rarely seen in males.(3) Ah-type ABNs were also identified by fluorescence.(4) Morphological data showed that myelinated fiber in ADN was ~25% of total and this result was consistent with our electrophysiological data.(5) Firing frequency of Ah-types(20~40 Hz) was lowered than that of A-types(40~150 Hz,P