Correlations of Myeloperoxidase (MPO), Adenosine deaminase (ADA), C–C motif chemokine 22 (CCL22), Tumour necrosis factor alpha (TNFα) and Interleukin-6 (IL-6) mRNA expression in the nasopharyngeal specimens with the diagnosis and severity of SARS-CoV-2 infections

Figures & data

Figure 1. Flowchart of patient inclusion in this study. Severe disease is defined as the need for supplemental oxygen on admission, admission to the intensive care unit (ICU) due to COVID-19, or death. Abbreviations: NPS, Nasopharyngeal specimens.

Table 1. Clinical characteristics and outcome of all patients included in the study * p < 0.05 ** < 50% of the uninfected patients had blood taken for lactate dehydrogenase and C-reactive protein. Data are expressed as n (%) or median (inter-quartile range), unless otherwise stated.

	Uninfected patients	Infected patients	p-value
Total number of cases	163	154
Age – median (Interquartile range)	85 (71–90)	82 (69–89)	0.191
Female	95 (58.3%)	77 (50.0%)	0.086
Number of deaths	7 (4.3%)	46 (29.9%)	<0.001*
Past Medical History
Hypertension	95 (58.3%)	84 (54.5%)	0.289
Diabetes Mellitus	54 (33.1%)	53 (34.4%)	0.451
Hyperlipidaemia	51 (31.3%)	43 (27.9%)	0.297
Cardiovascular disease	31 (19.0%)	39 (25.3%)	0.112
Chronic Renal Impairment	27 (16.6%)	41 (26.6%)	0.020*
Cerebral Vascular Accident	42 (25.8%)	35 (22.7%)	0.309
Obesity	2 (1.2%)	1 (0.6%)	0.521
Chronic Liver Disease	3 (1.8%)	13 (8.4%)	0.007*
Chronic Lung Disease	22 (13.5%)	21 (13.6%)	0.550
Solid Organ Malignancy	29 (17.8%)	18 (11.7%)	0.085
Haematological Malignancy	7 (4.3%)	10 (6.5%)	0.268
Solid Organ Transplantation	3 (1.8%)	9 (5.8%)	0.057
Haematological  Transplantation	1 (0.6%)	0 (0.0%)	0.514
Autoimmune Disease	1 (0.6%)	12 (7.8%)	0.001*
Immunosuppressant	18 (11.0%)	30 (19.5%)	0.026*
Clinical Presentation of infected patients
Fever	–	72 (46.8%)	–
Cough	–	68 (44.2%)	–
Sputum	–	40 (26.0%)	–
Shortness of breath	–	83 (53.9%)	–
Chest pain	–	9 (5.8%)	–
Gastrointestinal Symptoms	–	12 (7.8%)	–
Reasons for admission of uninfected patients
Infection	51 (31.3%)	–	–
Cardiovascular/ Pulmonary	25 (15.3%)	–	–
Gastrointestinal	8 (4.9%)	–	–
Malignancy	17 (10.4%)	–	–
Metabolic	6 (3.7%)	–	–
Neurological	14 (8.6%)	–	–
Trauma	23 (14.1%)	–	–
Clinical admission	19 (11.7%)	–	–
Blood Parameters on admission – median (interquartile range)
Haemoglobin, g/dL	10.4 (8.7–12.0)	11.4 (9.4–13.1)	0.005*
White Blood Cell, × 10^9/L	7.9 (6.1–10.5)	7.9 (5.1–11.9)	0.709
Neutrophil, × 10^9/L	5.6 (4.0–7.9)	6.3 (3.6–9.5)	0.249
Lymphocyte, × 10^9/L	1.1 (0.8–1.6)	0.8 (0.4–1.2)	<0.001*
Platelet, × 10^9/L	241 (179–313)	192 (137–279)	0.001*
Sodium, mmol/L	138 (134–140)	135 (132–140)	0.005*
Potassium, mmol/L	3.8 (3.6–4.3)	4.0 (3.6–4.4)	0.020*
Urea, mmol/L	6.2 (4.8–10.0)	8.8 (5.5–17.6)	<0.001*
Creatinine, mmol/L	82 (59–117)	107 (72–179)	<0.001*
Alkaline Phosphatase, U/L	83 (66–107)	83 (67–123)	0.494
Alanine Transaminase, U/L	16 (11–27)	25 (16–41)	<0.001*
Lactate Dehydrogenase**, U/L	252 (202–293)	348 (256–497)	<0.001*
C-reactive protein**, mg/dL	1.9 (0.8–7.8)	6.5 (1.8–11.9)	0.001*
Vaccination Status			<0.001*
Complete	93 (57.1%)	42 (27.3%)
Incomplete/Unvaccinated	70 (42.9%)	112 (72.7%)

Figure 2. Comparison of normalized and total MPO, ADA, CCL22, TNFα, and IL-6 mRNA expression in nasopharyngeal specimens between COVID-19 infected and uninfected individuals. The bar represents the median of the samples, the hinges mark the interquartile range. *p < 0.05, **p < 0.005, ***p < 0.001.

Figure 3. Receiver operating characteristics (ROC) curve of normalized MPO, ADA, CCL22, TNFα, and IL-6 mRNA expression in identification of COVID-19 infected patients. The dotted line represents the line of identity.

Figure 4. Comparison of normalized MPO, ADA, CCL22, TNFα, and IL-6 mRNA expression in nasopharyngeal swabs between different specimens in COVID-19 patients. The central point represents the median of the samples, the hinges mark the interquartile range. *p < 0.05, **p < 0.005, ***p < 0.001.

Table 2. Clinical characteristics and outcome of all COVID-19 infected patients classified according to severity of disease. Data are expressed as n (%) or median (inter-quartile range), unless otherwise stated. * p < 0.05.

	Mild cases	Severe cases	p-value
Total number of cases	59	95
Age – median (Interquartile range)	75 (64–87)	83 (72–90)	0.029*
Female	32 (54.2%)	45 (47.4%)	0.254
Ct value of first NPS specimen	26.5 (20.8–32.3)	25.7 (22.0–29.9)	0.617
Past Medical History
Hypertension	29 (49.2%)	55 (57.9%)	0.186
Diabetes Mellitus	20 (33.9%)	33 (34.7%)	0.529
Hyperlipidaemia	16 (27.1%)	27 (28.4%)	0.506
Ischaemic Heart Disease	15 (25.5%)	24 (25.3%)	0.564
Chronic Renal Impairment	17 (28.8%)	24 (25.3%)	0.381
Cerebral Vascular Accident	12 (20.3%)	23 (24.2%)	0.363
Obesity	0 (0.0%)	1 (1.1%)	0.617
Chronic Liver Disease	6 (10.2%)	7 (7.4%)	0.372
Chronic Lung Disease	4 (6.8%)	17 (17.9%)	0.040*
Solid Organ Malignancy	9 (15.3%)	9 (9.5%)	0.203
Haematological Malignancy	2 (3.4%)	8 (8.4%)	0.187
Solid Organ Transplantation	3 (5.1%)	6 (6.3%)	0.525
Haematological  Transplantation	0 (0.0%)	0 (0.0%)	–
Autoimmune Disease	4 (6.8%)	8 (8.4%)	0.485
Immunosuppressant	11 (18.6%)	19 (20.0%)	0.505
Clinical Presentation
Fever	26 (44.1%)	46 (48.4%)	0.360
Cough	20 (33.9%)	48 (50.5%)	0.031*
Sputum	9 (15.3%)	31 (32.6%)	0.012*
Shortness of breath	15 (25.4%)	68 (71.6%)	<0.001*
Chest pain	6 (10.2%)	3 (3.2%)	0.075
Gastrointestinal Symptoms	5 (8.5%)	7 (7.4%)	0.515
Blood Parameters on admission – median (interquartile range)
Haemoglobin, g/dL	10.6 (8.9–13.1)	11.5 (9.5–13.1)	0.292
White Blood Cell, × 10^9/L	6.0 (3.9–9.4)	8.4 (6.2–12.9)	0.003*
Neutrophil, × 10^9/L	4.3 (3.0–7.6)	7.0 (4.5–10.8)	0.001*
Lymphocyte, × 10^9/L	0.9 (0.5–1.3)	0.8 (0.4–1.2)	0.451
Platelet, × 10^9/L	201 (129–285)	191 (143–260)	0.707
Sodium, mmol/L	135 (132–139)	136 (132–142)	0.172
Potassium, mmol/L	3.9 (3.5–4.3)	4.1 (3.6–4.6)	0.083
Urea, mmol/L	7.6 (4.7–11.3)	10.2 (5.8–23.0)	0.021*
Creatinine, mmol/L	91 (69–139)	120 (77–211)	0.014*
Alkaline Phosphatase, U/L	75 (66–124)	89 (67–123)	0.319
Alanine Transaminase, U/L	23 (15–40)	26 (17–45)	0.371
Lactate Dehydrogenase, U/L	258 (205–364)	417 (283–558)	<0.001*
C-reactive protein, mg/dL	4.9 (0.6–8.8)	7.4 (2.5–14.2)	0.006*
Vaccination Status			0.009*
Complete	23 (39.0%)	19 (20.0%)
Incomplete/Unvaccinated	36 (61.0%)	76 (80.0%)

Figure 5. Comparison of normalized MPO, ADA, CCL22, TNFα, and IL-6 mRNA expression in nasopharyngeal swabs between different specimens in patients with mild and severe COVID-19. The central point represents the median of the samples, the hinges mark the interquartile range. *p < 0.05, **p < 0.005, ***p < 0.001.

Table 3. Multivariate analysis of severe COVID-19 Infection. Mean of CCL22/GAPDH from each patient was obtained, and logarithmic transformation of the ratio was performed for easy interpretation of odds ratio generated by logistic regression. * p < 0.05.

Parameters	Multivariate model OR	p-value
Age	–	–
Log (Average CCL22/ GAPDH)	0.211 (0.060–0.746)	0.016*
Chronic Lung Disease	7.771 (0.593–101.872)	0.118
Leucocytosis (WCC > 9.93 × 10^9/L)	–	–
Neutrophilia (ANC > 7.42 × 10^9/L)	3.220 (0.883–11.736)	0.076
Urea > 4.4 mmol/L	–	–
Creatinine > 83 umol/L	–	–
LDH > 221 U/L	10.146 (2.275–45.252)	0.002*
CRP > 0.76 mg/dL	–	–
Cough	4.617 (1.230–17.328)	0.023*
Sputum	4.465 (0.710–28.094)	0.111
Shortness of breath	13.742 (3.938–47.962)	<0.001*
Completed vaccination	0.167 (0.035–0.794)	0.024*

Data availability statement

The data that support the findings of this study are available from the corresponding author, K.Y. Yuen, upon reasonable request.