Metagenomic Next-Generation Sequencing for Diagnostically Challenging Mucormycosis in Patients with Hematological Malignancies

Figures & data

Table 1 Characteristics of Patients

	Characteristics	Number (%)
Gender	Male	9 (69.23)
	Female	4 (30.77)
Age	Median age	43 (14–73)
Basic diagnosis	AML	3 (23.08)
	ALL	5 (38.46)
	MDS	3 (23.08)
	HAL	1 (7.69)
	ITP	1 (7.69)
Primary disease state	CR	10 (76.92)
	NR	3 (23.08)
Current medication	Immunosuppression or Glucocorticoids	6 (46.15)
	Chemotherapy within 3 months	7 (53.85)
Neutropenia	<500/µL	8 (61.54)
Previous treatment	Allo-HSCT	3 (23.08)
	CAR-T	1 (7.69)
	CAR-T+Allo-HSCT	3 (23.08)
	Chemotherapy alone	6 (46.15)
Mucormycosis diagnosis	Proven	4 (30.77)
	Probable	9 (69.23)

Abbreviations: AML, Acute myelocytic leukemia; ALL, Acute lymphocytic leukemia; MDS, Myelodysplastic syndromes; HAL, hybrid acute leukemia; ITP, Idiopathic thrombocytopenic purpura; CR, complete remission; NR, no remission; Allo-HSCT, Allogeneic hematopoietic stem cell transplantation; CAR-T, Chimeric Antigen Receptor T-Cell Immunotherapy.

Table 2 Characteristics of “Proven” Patients

Num	Sex/Age	Symptom/Date of First Clinical Symptoms	mNGS/Date	G	GM	Infectionsites/Histology/Date	Antifungal	Surgery	Outcome at Day 90	Days From mNGS to Histology
1	M/37	Fever, Hemoptysis/D-8	Mucor/D3	N	N	Pulmonary/Mucor/D45	L-AmB, and posaconazole	No	Death/D49	D42
2	M/43	Fever, pectoralgia,Hemoptysis/D-7	Rhizomucor pusillus/D5	N	N	Pulmonary/Rhizomucor pusillus/D2	L-AmB and posaconazole	No	Alive	D2
3	M/52	Fever, Cough/D-1	Rhizomucor pusillus/D15	N	N	Pulmonary/Muco/D17	L-AmB and posaconazole	No	Alive	D2
4	F/27	Headache/D-4	Rhizopus oryzae/D2	N	N	Sinus/Rhizopus oryzae/D6	L-AmB	Yes	Alive	D4

Abbreviations: G, 1,3 beta-Dglucan detection test; GM, galactomannan; L-AmB, liposomal amphotericin B; mNGS, Metagenomic next-generation sequencing; N, negative.

Table 3 Characteristics of “Probable” Patients

Num	Sex/Age	Symptom/Date of First Clinical Symptoms	mNGS/Date	G	GM	Infectionsites/Histology/Date	Antifungal	Surgery	Outcome at Day 90
5	F/42	Fever, Coma/D-2	Rhizopus oryzae/D-2	N	N	Cerebral/-/-	L-AmB	No	Death/D4
6	M/73	Fever/D-3	Rhizomucor pusillus/D4	N	N	Pulmonary/-/-	L-AmB	No	Death/D30
7	M/58	Fever/D-19	Rhizopus oryzae/D4	N	N	Pulmonary/-/-	L-AmB, and posaconazole	No	Alive
8	F/14	Fever, Swelling of right eye, Blurred vision, Headache/D-7	Rhizopus oryzae/D2	N	N	Rhino-cerebral/-/-	L-AmB/D2	No	Death/D2
9	M/40	Headache/D-7	Mucor/D3	N	N	-	L-AmB, and posaconazole	Yes	Alive
10	M/38	Fever/D-5	Rhizopus oryzae/D7	118.86	N	Pulmonary/-/-	L-AmB, and posaconazole	No	Death/D70
11	M/64	Fever/D-1	Rhizomucor pusillus/D0	N	N	Pulmonary/-/-	L-AmB, and posaconazole	No	Death/D12
12	M/48	Fever/D-2	Rhizopus microspores/D1	267.57	N	Pulmonary/-/-	-	No	Death/D2
13	F/59	Pectoralgia/D-3	Rhizopus microspores/D2	N	N	Pulmonary/-/-	L-AmB, and Posaconazole	No	Alive

Abbreviations: G, 1,3 beta-Dglucan detection test; GM, galactomannan; L-AmB, liposomal amphotericin B; mNGS, Metagenomic next-generation sequencing; N, negative.

Figure 1 The distribution of mNGS sample and infection site. (A) Distribution of mNGS samples in the “Proven” and “Probable” group. (B) The infection site in the “Proven” and “Probable” group.

Figure 2 Imaging results of patients 1 to 4. The imaging results of patients 1–4 indicate that there may be fungal infection. (A) Chest CT indicates right pneumonia and pneumothorax. (B) Chest CT indicates bilateral pulmonary inflammation and bilateral pleural effusion. (C) Chest CT shows nodules in the anterior basal segment of the lower lobe of the left lung. (D) Imaging results showed inflammation of the right maxillary sinus and sphenoid sinus.

Figure 3 Imaging changes of patient 4. (A) Imaging results in D0 showed inflammation of the right maxillary sinus and sphenoid sinus, suggesting possible fungal infection. (B) The imaging results before surgical debridement (D5) indicated that the inflammation had progressed; (C) Imaging results after treatment (D16) suggest that inflammation is relieved. Arrows indicate the site of infection.