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Background: The imported cases of Plasmodium malariae (P. malariae) and Plasmodium ovale (P. ovale) malaria are increasing annually, especially in central China. Here, we report a case of serological rapid diagnostic test (RDT)-negative P. malariae malaria imported from West Africa., Methods: The case patient was exclusively diagnosed with P. malariae through microscopy, Plasmodium genus-specific nested polymerase chain reaction (PCR), and sequencing of targeted P. malariae circumsporozoite (pmcsp) gene, except for serological RDT., Results: The patient was discharged in stable condition after 5 days of hospitalization, with no overt malaria parasites or associated symptoms., Conclusions: This case reveals that asymptomatic P. malariae infections can occur among exported laborers back from malaria-endemic areas, some of whom may escape serological screening test or RDT, posing a continuing potential threat to malaria control. Therefore, PCR-based molecular techniques are more effective and necessary than serological RDT for malaria surveillance nationwide.