Diagnostic microbiology and infectious disease · 2025
Diagnostic performance of an in-house real-time polymerase chain reaction (PCR) assay and antigen detection from respiratory samples for the diagnosis of pulmonary cryptococcosis.
СтатьяОценочное исследованиеНазвание и аннотация: Перевод в очереди
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Kumar Karthick, Choudhary Hansraj, Ahmad Haseen, Muthu Valliappan, Gupta Parikshaa, Rudramurthy Shivaprakash M, Agnihotri Sourav, Agarwal Ritesh, Chakrabarti Arunaloke, Kaur Harsimran
Аннотация
INTRODUCTION: The currently available diagnostic tests lack sensitivity to diagnose pulmonary cryptococcosis. In the current study, we developed and standardized an in-house real-time PCR assay and evaluated the antigen detection in respiratory samples for the diagnosis of pulmonary cryptococcosis.
MATERIALS AND METHODS: We standardized an in-house real-time PCR assay (using URA5 and STR1 primers; index test 1) and cryptococcal antigen detection (BIOSYNEX® CryptoPS, France) from the respiratory samples (index test 2). We considered a sample positive for PCR assay when both gene targets (URA5 and STR1) were detected. We prospectively enrolled subjects undergoing evaluation for non-resolving pneumonia and evaluated the performance of the index tests for diagnosing pulmonary cryptococcosis. The reference standard was proven or probable pulmonary cryptococcosis diagnosed by EORTC/MSG (European Organization for Research and Treatment of Cancer/ Mycoses Study Group) criteria.
RESULTS: Of the 133 subjects enrolled in the study, two (2.66 %) and three (3.99 %) were diagnosed as having proven and probable pulmonary cryptococcosis. 3.8 % of study subjects had HIV. The sensitivity and specificity of qPCR (index test 1) for the diagnosis of pulmonary cryptococcosis were 60.0 % (95 % CI: 14.6-94.7) and 96.1 % (95 % CI: 91.1-98.7), respectively, while the sensitivity and specificity of antigen detection from respiratory samples (index test 2) were 40.0 % (95 % CI: 5.2-85.3) and 99.2 % (95 % CI: 95.7 -100.0), respectively.
DISCUSSION: In-house PCR and antigen detection in respiratory specimens can potentially be used for early diagnosis of pulmonary cryptococcosis. Larger multicenter studies are required to confirm their utility.
MATERIALS AND METHODS: We standardized an in-house real-time PCR assay (using URA5 and STR1 primers; index test 1) and cryptococcal antigen detection (BIOSYNEX® CryptoPS, France) from the respiratory samples (index test 2). We considered a sample positive for PCR assay when both gene targets (URA5 and STR1) were detected. We prospectively enrolled subjects undergoing evaluation for non-resolving pneumonia and evaluated the performance of the index tests for diagnosing pulmonary cryptococcosis. The reference standard was proven or probable pulmonary cryptococcosis diagnosed by EORTC/MSG (European Organization for Research and Treatment of Cancer/ Mycoses Study Group) criteria.
RESULTS: Of the 133 subjects enrolled in the study, two (2.66 %) and three (3.99 %) were diagnosed as having proven and probable pulmonary cryptococcosis. 3.8 % of study subjects had HIV. The sensitivity and specificity of qPCR (index test 1) for the diagnosis of pulmonary cryptococcosis were 60.0 % (95 % CI: 14.6-94.7) and 96.1 % (95 % CI: 91.1-98.7), respectively, while the sensitivity and specificity of antigen detection from respiratory samples (index test 2) were 40.0 % (95 % CI: 5.2-85.3) and 99.2 % (95 % CI: 95.7 -100.0), respectively.
DISCUSSION: In-house PCR and antigen detection in respiratory specimens can potentially be used for early diagnosis of pulmonary cryptococcosis. Larger multicenter studies are required to confirm their utility.