The invention relates to a kit for simultaneous detection of Anopheles moltatus, Anopheles reckeii and Anopheles lanceolata by multiplex PCR. The kit includes specific primers for Anopheles papillonus, Anopheles reckeii, and Anopheles beriorhynchus, 10×buffer, dNTPs, MgCl 2 , Taq enzyme, template DNA, sterile double-distilled water, the sequences of the specific primers of Anopheles moltatus, Anopheles reckeii, and Anopheles lanceolata are respectively as SEQ ID NO.5-6, SEQ ID NO.15 -16, shown in SEQ ID NO.33-34. The multiplex PCR detection kit of the present invention can give qualitative detection results within 3 hours after receiving the DNA sample, saves cost and time, and has low requirements for personnel and equipment. and an effective tool for An.