3.2. Study Population and Eligibility Criteria
A total of 104 healthy children aged 1 month to 2 years who presented to the Social Pediatrics outpatient clinic of Kütahya Health Sciences University Hospital for routine checkups and had no known underlying diseases at the time of evaluation were included as the control group. These individuals were matched to the patient group by age and sex, and their laboratory parameters were retrospectively retrieved from the hospital information system.
Bronchiolitis was diagnosed clinically according to the American Academy of Pediatrics (AAP) criteria (
12). In infants aged 1 - 24 months, the diagnosis required at least 2 clinical features, including an increased respiratory rate (tachypnea), intercostal or subcostal chest wall retractions, and abnormal lung sounds, such as wheezing or crackles on auscultation (
12-
15) (
Table 1).
| Parameter | Mild | Moderate | Severe |
|---|
| Feeding | Normal | Less than usual; can take more than 50% of usual daily oral intake | Not interested; can take less than 50% of usual daily oral intake |
| Apnea | No | No | Yes |
| Respiratory rate/min | < 50 | 50 - 70 | > 70 |
| Heart rate, beats/min | < 140 | 140 - 160 | > 160 |
| Retractions | Mild | Moderate | Severe |
| Cyanosis | Absent | Absent | Present |
| Oxygen saturation (%) | > 92 | 90 - 92 | < 90 |
| Respiratory support | None | HFNC/NIV | IMV or failure of non-invasive support |
| Clinical course | Stable | May require support | Deterioration/progression |
| Complications | Absent | Absent | Present, such as ARDS |
a Abbreviations: ARDS, acute respiratory distress syndrome; HFNC, high-flow nasal cannula; IMV, invasive mechanical ventilation; NIV, non-invasive ventilation.
Bronchiolitis severity was classified as moderate or severe based on predefined clinical criteria and the level of respiratory support required. In our tertiary intensive care unit, respiratory support is initiated according to the patient’s clinical status at presentation. Non-invasive modalities, including high-flow nasal cannula (HFNC) or non-invasive ventilation (NIV), are used as first-line therapy, with escalation to invasive mechanical ventilation (IMV) in cases of insufficient clinical response or clinical deterioration.
For the purposes of this study, severity classification was operationalized according to the highest level of respiratory support required during hospitalization, together with key clinical findings. Severe bronchiolitis was defined as the requirement for IMV at any time during the clinical course or the presence of at least one of the following: oxygen saturation < 90% on room air requiring continuous oxygen supplementation or signs of severe respiratory distress, including marked retractions, grunting, or apnea. Moderate bronchiolitis was defined as respiratory symptoms requiring hospitalization and supplemental oxygen, managed with HFNC or NIV without progression to IMV.
Patients were classified according to the most severe criterion met during their clinical course. Bronchiolitis severity was defined using a composite framework integrating clinical findings, respiratory support requirements, and disease course, as summarized in
Table 1. This approach reflects real-world clinical practice and minimizes subjectivity in severity classification. Severity classification was performed retrospectively through a structured review of electronic medical records by the study investigators. In cases with multiple criteria, classification was based on the highest level of severity observed during hospitalization. All severity assessments were performed by 2 pediatric intensive care physicians, and discrepancies were resolved by consensus.
The study included patients younger than 2 years who were hospitalized and diagnosed with AB after clinical and laboratory evaluation by a pediatrician.
Participants were categorized into 2 groups: individuals with AB and healthy control subjects. Children with AB were further subdivided into moderate and severe bronchiolitis groups. The hematological, biochemical, and inflammatory indices of each patient were examined. These included hemogram parameters, including leukocytes (white blood cell [WBC] count), hemoglobin (Hb), and platelet count; biochemical parameters, including C-reactive protein (CRP), procalcitonin (PCT; ng/mL), and albumin; and the HALP score.
Data collected included age at admission, sex, past medical history, clinical presentation, and vital signs (heart rate and respiratory rate). For each patient, the clinical severity of AB, duration of PICU stay, need for respiratory support, high-flow oxygen therapy, and invasive and non-invasive forms of mechanical ventilation, along with their durations, were recorded.
The HALP score was calculated as follows: hemoglobin (g/L) × albumin (g/L) × lymphocyte count (cells/L) ÷ platelet count (cells/L) (
16). For score computation, laboratory values obtained at the patient’s initial presentation to the pediatric emergency department, before intravenous access, hydration, or initiation of any systemic therapy, were used. When multiple blood samples were available, only the earliest measurement was included in the analysis.
Patients with severe immunodeficiency, prematurity, chronic lung disease (cystic fibrosis, bronchopulmonary dysplasia, etc), cardiac disease (congestive heart failure, cyanotic congenital heart disease, etc), and hematological diseases that may affect lung function and HALP score parameters, such as neurometabolic diseases, were excluded from the study population. Participants receiving treatment at an external healthcare facility, including patients receiving inhaled steroids/bronchodilators, systemic steroids, or intravenous hydration, were also excluded. This study involved analysis of data from children with moderate to severe AB admitted to a tertiary PICU.
A total of 140 children diagnosed with AB were initially screened. Of these, 40 patients were excluded based on predefined criteria, including prematurity or bronchopulmonary dysplasia (n = 18), prior inpatient treatment at another center (n = 10), neurometabolic disease (n = 6), suspected immunodeficiency (n = 3), dilated cardiomyopathy (n = 2), and heart failure (n = 1). Consequently, 100 patients met the inclusion criteria and were included in the final analysis. The participant flow is presented in
Figure 1.
Flowchart of patients included in the study
Clinical outcomes were predefined and extracted from electronic medical records for the index hospitalization. All time-based outcomes were measured in days. Total hospital stay was defined as the number of days from hospital admission to discharge. PICU stay was defined as the duration of admission to the PICU during the same hospitalization. Time to transition to room air was defined as the number of days from admission until discontinuation of supplemental oxygen support.
Duration of non-invasive ventilation was defined as the total number of days during which the patient received HFNC or NIV. Duration of IMV was defined as the total number of days on invasive ventilatory support. Total duration of respiratory support was defined as the cumulative duration of all respiratory support modalities. All outcomes were calculated based on the index hospitalization. Patients with incomplete outcome data, transfers to other institutions, or missing follow-up information were excluded from outcome analyses. Because length-of-stay and duration variables showed a non-normal distribution, non-parametric methods were used for group comparisons.
3.3. Statistical Analysis
Statistical analyses were performed using SPSS version 28.0 (IBM Corp., Armonk, NY, USA). Categorical variables were presented as frequencies and percentages, whereas continuous variables were summarized as mean ± standard deviation or median (minimum-maximum), as appropriate. The Kolmogorov-Smirnov test was used to assess the normality of the data distribution. Comparisons between groups were performed using the independent-samples t-test for normally distributed variables and the Mann-Whitney U test for non-normally distributed variables. Categorical variables were compared using the chi-square test.
Given the potential influence of confounding factors on HALP components, including age, sex, inflammatory burden, nutritional status, and possible bacterial coinfection, these variables were considered during analysis and interpretation. In particular, age and sex were evaluated descriptively, and inflammatory markers, including CRP and PCT, were examined to contextualize the findings. However, because of the retrospective design and relatively limited sample size, comprehensive multivariable adjustment for all potential confounders was not feasible; therefore, the results were interpreted with caution, and the possibility of residual confounding was acknowledged.
In addition to P values, effect sizes were considered to improve the clinical interpretability of between-group differences, and median differences were calculated for key variables. Because multiple laboratory parameters were evaluated, the potential for type I error due to multiple comparisons was recognized. Formal adjustment methods were not applied because of the exploratory nature of the study; accordingly, findings were interpreted conservatively. Where applicable, the number of observations (n) for each laboratory parameter was reported to account for missing data.
Receiver operating characteristic (ROC) curve analysis was performed to evaluate the discriminative performance of the HALP score. The optimal cut-off value was determined using the Youden Index. Sensitivity and specificity corresponding to the optimal cut-off were calculated. The area under the curve (AUC) was reported with 95% confidence intervals (CIs) to quantify overall diagnostic accuracy. The binary outcomes for ROC analyses were defined as AB versus control for diagnostic performance and severe versus non-severe bronchiolitis for severity assessment. All ROC analyses were conducted on the same dataset used to derive the cut-off values; therefore, the potential for optimism bias was acknowledged. Internal validation methods, such as bootstrapping or cross-validation, were considered; however, when not feasible, the risk of overfitting was explicitly acknowledged, and findings were interpreted conservatively. A P value < 0.05 was considered statistically significant. Because of the relatively limited sample size and the number of potential confounding variables, multivariable regression analysis was not performed. Therefore, no adjusted estimates are presented, and the findings should be interpreted as exploratory. Potential confounding effects, particularly those related to age and sex, could not be fully controlled.