The chloroform fumigation extraction (CFE) method determines microbial biomass carbon (MBC) or nitrogen (MBN) by calculating the increase in extractable carbon (C) or nitrogen (N) due to microbial lysis during chloroform fumigation. In China, many studies have focused on the impacts of N and phosphorus (P) addition on soil MBC and MBN in forest ecosystems, where substantial atmospheric N deposition has strongly acidified soils. The addition of nutrients may alter the extraction process applied in the CFE method, potentially influencing the MBC and MBN determined by the CFE method independently of the actual microbial biomass. In this study, we tested whether the MBC and MBN determined by the CFE method were biased by the experimental addition of N and P in strongly acidified Chinese forest soils by adding N and P to the soils immediately before chloroform fumigation, which should not affect the actual microbial biomass. P addition significantly elevated the dissolved organic carbon (DOC) content, especially after fumigation, while N addition significantly reduced the dissolved nitrogen (DN) content. The added N was subtracted using blank samples without soil. However, the altered DOC and DN contents did not affect the MBC and MBN contents determined by the CFE method. In conclusion, our study suggests that the CFE is a relatively robust method to test the impacts of nutrient addition on microbial biomass in the strongly acidified soils of Chinese forests. We also suggested that: (i) even if a fertilization experiment results in an elevated DOC content following P addition, it does not necessarily indicate a stimulation of DOC production by microbes; and (ii) the soil adsorption capacity or the strength of microbial N uptake during the extraction procedure applied in the CFE method may affect the determination of MBN by influencing the DN extraction efficiency.