Meloidogyne enterolobii Yang & Eisenback, 1983 (guava root-knot nematode) is an important disease in several subtropical to tropical areas of the world (Subbotin et al. 2021). It is a highly polyphagous root-knot nematode species causing major damage to a range of economically important crops. The expansion of this species is increasing worldwide, creating a potential problem to the maintenance of resistance genes to other major Meloidogyne species (Castagnone-Sereno and Castillo 2020). Diagnosis of M. enterolobii can be challenging due to morphological similarities with other root-knot nematode species (Castagnone-Sereno 2012). It has been reported in several countries of Equatorial and South Africa (Subbotin et al. 2021), but not in North Africa. Two guava groves in Egypt (Bany Salama, Natrn valley, El Beheira governorate, 30.322043N, 30.518529E; and Izbat Al Halawijah, Monshaah Alaweyah, Abu El Matamir, El Beheira governorate 30.9398050N, 30.1484430E) were found with significant symptoms of tree decline and root galling damage. Egg masses and female root-knot nematodes were found inside the galls. Nematodes were extracted from soil samples with levels of 12,300 and 12,600 second-stage juveniles (J2s)/250 g of soil using a modified Baerman method (Hooper 1986), respectively. Nematode root density was 24.367 eggs/g of root, using the protocol of Hussey and Barker (1973), for the Izbat Al Halawijah population. For morphological and morphometrical identification, J2s and females were fixed using a hot formalin solution (4% v/v). DNA was isolated from single J2 specimens for: (i) testing multiplex specific-PCR assay for M. incognita, M. javanica, and M. arenaria (Kiewnick et al. 2013), and (ii) amplifying and sequencing of cytochrome oxidase subunit II (COII) and the 16S rRNA mitochondrial region using the primer pair C2F3 (5′-GGTCAATGTTCAGAAATTTGTGG-3′) (Powers and Harris 1993) and MRH106 (5′-AATTTCTAAAGACTTTTCTTAGT-3′) (Stanton et al. 1997). Perineal patterns of females from Izbat Al Halawijah were typical of the species, body size (L: 520 to 774 μm; W: 214 to 487 μm), stylet length (12.5 to 13.7 μm), and ratio of distance from anterior end to excretory pore and stylet length (4.2) in females (n = 18), fit the original description and others (Subbotin et al. 2021). J2s from Izbat Al Halawijah (n = 13) showed: body length (393.5 to 475 μm), stylet length (11.5 to 13.5 μm), excretory pore to anterior end (89 to 95.5 μm), tail length (50.0 to 60.0 μm), tail hyaline region (12.0 to 21.0 μm), a ratio (24.2 to 32.5), b ratio (4.9 to 6.5), c ratio (7.3 to 8.6), and c’ (5.0 to 6.4), also fitting with the original description and others (Subbotin et al. 2021). Specific PCR did not amplify any band (Kiewnick et al. 2013). Four J2s were sequenced for COII-16S rRNA region for each population showing M. enterolobii as a unique species and without intraspecific variability. Two identical DNA fragments of 814 bp obtained for both populations (OP434400 and OP434401) were compared with those in GenBank. A BLAST search indicated the sequences were 100% identical to several sequences of M. enterolobii (MF467278 and KX823371). Based on these results, the root-knot nematodes isolated from these two guava groves in Egypt were confirmed as M. enterolobii. This is a well-known pathogen of guava, causing important losses in this crop (Castagnone-Sereno and Castillo 2020), and it is regulated as a quarantine nematode in the Mediterranean region (EPPO).