Upadhyay Pallavi, Vallabhaneni Arjuna, Ager Edward, Alexander Barbara, Rosato Adriana, Singh Vijay
HealthTrackRx, 1500 I-35 W, Denton, TX 76207, USA.
Division of Infectious Diseases, Departments of Medicine and Pathology, Duke University, Durham, NC 27701, USA.
Diagnostics (Basel). 2025 Aug 1;15(15):1939. doi: 10.3390/diagnostics15151939.
: Unambiguous clinical interpretation of PCR results for urinary tract infections (UTIs) remains a challenge. Here we compare and correlate multiplex qPCR results (quantification cycle values) with traditional microbial culture results (colony forming units) for clinical samples. : Serial dilutions [10 to 10 colony forming units (CFU)/mL] were performed on five Gram-negative and two Gram-positive UTI-causing bacterial pathogens. For each dilution, quantitative cultures on solid media to confirm CFU/mL values and a real-time PCR UTI panel employing a nanofluidic Open Array platform producing quantification cycle (Cq) values were performed. Cq values were correlated with CFU/mL values, generating a semi-quantitative interpretive scale for clinical samples. The clinical utility of the scale was then assessed using PCR and culture data from 168 clinical urine samples. : For Gram-negative bacteria, Cq values of <23, 23 to 28, and >28 corresponded with ≥10 CFU/mL, <10 CFU/mL and negative cultures, respectively. For Gram-positive bacteria, Cq values of <26, 26 to 30, and >30 corresponded with ≥10 CFU/mL, <10 CFU/mL and negative cultures, respectively. Among 168 urine specimens (including 138 Gram-negative and 30 Gram-positive bacteria), there was 83.3% agreement ( = 140/168) and 16.6% non-agreement ( = 28/168) between culture CFU/mL and qPCR Cq. Gram-negative bacteria had higher agreement (87.6%, 121/138) than Gram-positive bacteria (63.3%, 19/30). : This study demonstrates that qPCR Cq results can be directly correlated with traditional urine quantitative culture results and reliably identify the clinically relevant cutoff of 10 CFU/mL for detected uropathogens.
对尿路感染(UTIs)的聚合酶链反应(PCR)结果进行明确的临床解读仍然是一项挑战。在此,我们将临床样本的多重定量聚合酶链反应(qPCR)结果(定量循环值)与传统微生物培养结果(菌落形成单位)进行比较并关联。:对五种革兰氏阴性和两种革兰氏阳性引起UTIs的细菌病原体进行系列稀释[10至10菌落形成单位(CFU)/mL]。对于每种稀释度,在固体培养基上进行定量培养以确认CFU/mL值,并使用产生定量循环(Cq)值的纳米流体开放式阵列平台进行实时PCR UTI检测。将Cq值与CFU/mL值相关联,为临床样本生成半定量解释标准。然后使用来自168份临床尿液样本的PCR和培养数据评估该标准的临床实用性。:对于革兰氏阴性菌,Cq值<23、23至28和>28分别对应≥10 CFU/mL、<10 CFU/mL和培养阴性。对于革兰氏阳性菌,Cq值<26、26至30和>30分别对应≥10 CFU/mL、<10 CFU/mL和培养阴性。在168份尿液标本(包括138份革兰氏阴性菌和30份革兰氏阳性菌)中,培养CFU/mL与qPCR Cq之间的一致性为83.3%(=140/168),不一致性为16.6%(=28/168)。革兰氏阴性菌的一致性(87.6%,121/138)高于革兰氏阳性菌(63.3%,19/30)。:本研究表明,qPCR Cq结果可直接与传统尿液定量培养结果相关联,并可靠地确定检测到的尿路病原体的临床相关临界值为10 CFU/mL。