Table 2.

Advantages and disadvantages of different neutralizing antibody detection methods.

Method Classification	Method Name	Advantages	Disadvantages
cVNT	CPE	•The “gold standard” for neutralizing antibody detection has specificity, sensitivity, and can accurately measure the neutralizing ability of antibodies [76].	•BSL-3, low throughput, time-consuming, high condition requirements, and is influenced by subjective explanations from researchers, resulting in cumbersome steps [76,122].
	PRNT
	Recombinant live virus	•Reduced time consumption, dynamic monitoring, and objective results [70]. •Can be safely used at BSL-2 laboratories [84,85].	•Cumbersome and time-consuming steps [84,85].
pVNT	HIV system	•BSL-2, high throughput, less time-consuming, specific, sensitive, consistent with live virus results, accurately measure the neutralizing ability of antibodies [66]; •Lentiviral vector-based pseudoviruses differ from conventional retroviral vectors in that they have the ability to infect both dividing and non-dividing cells; •VSV systems are suitable for neutralizing antibody analysis or screening of high-content therapies [123].	•The number of envelope proteins in pseudoviruses is not directly proportional to the copy number of the core genome. The titer needs to be measured through qPCR and other methods, which loses its authenticity, especially when comparing the impact of different mutant S proteins on viral infectivity [105]; •VSV residue in the VSV system may lead to false positive results [111]; •Comparison and validation with live viruses are required.
	VSV system
	MLV system
sVNT	ELISA	•Cell-independent, safe; high throughput, low cost, less time-consuming [119]; can be used for screening for neutralizing antibodies in the population to investigate the efficacy of protective immunity and vaccination [124].	•Does not represent actual neutralizing antibody titers, does not provide information on the neutralizing ability of antibodies and has low sensitivity for early infection [67,125].