体外培养法与RNA等温扩增法对脲原体检测能力比较
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摘要
目的比较体外培养法与RNA等温扩增法对脲原体的检测能力。方法 2016年1-8月共收集了103份就诊于北京协和医院门诊患者的首次尿标本。标本平均分成3份,分别用于脲原体的检测。第1份采用RNA等温扩增法,第2份采用体外培养法,第3份采用体外培养联合测序法。其中,体外培养联合测序法将作为参考方法,横向评估体外培养法和RNA恒温扩增法对脲原体的检测能力。结果基于体外培养联合测序法,共有24例标本(23.30%)检测出了脲原体。其中,14例(17.50%)来自于男性,10例(43.47%)来自于女性,差异有统计学意义(P=0.022);40～<50岁脲原体检出率最高(5/12,41.67%)。与参考方法相比,体外培养法的灵敏度和特异度分别为75.00%(18/24)和100.00%(79/79);RNA等温扩增法的灵敏度和特异度分别为95.83%(23/24)和96.20%(76/79),差异无统计学意义(P>0.05)。在24份检测出脲原体阳性标本中,17例(70.83%)为微小脲原体,7例(29.17%)为解脲脲原体,未检查到微小脲原体和解脲脲原体共同存在的情况。18例培养阳性标本进行了体外药敏测定,其中环丙沙星和氧氟沙星的体外敏感性最差。结论与培养联合测序法相比,体外培养法和RNA等温扩增法均表现出了较好的灵敏度和特异度,但从临床应用角度来讲,二者联合使用可能会对脲原体感染的诊断和治疗提供更全面的指导。
Objective To compare the detection of Ureaplasma determined by in vitro culture and RNA isothermal amplification.Methods From January to August 2016,a total of 103 urine specimens were collected from outpatients of Peking Union Medical College Hospital.The specimens were divided into three parts for the detection of Ureaplasma.The first part was determined by RNA isothermal amplification assay,the second part was tested by in vitro culture,and the third part was detected by in vitro culture and sequencing.In vitro culture combined with sequencing will be used as a reference method to evaluate the detection ability of Ureaplasma tested by in vitro culture and RNA thermostatic amplification.Results Ureaplasma was detected in24 specimens(23.30%)based on in vitro culture and sequencing.Among them,14 specimens(17.50%)were male and 10 specimens(43.47%)were female,the difference was statistically significant(P=0.022).From the age group,the highest Ureaplasma detection rate was found in 40-<50 age group(5/12,41.67%).Compared with the reference method,the sensitivity and specificity of in vitro culture were 75%(18/24)and100.00%(79/79),and the sensitivity and specificity of RNA isothermal amplification were 95.83%(23/24)and 96.20%(76/79),respectively,the difference was not statistically significant(P>0.05).Of the 24 samples with positive ureaplasma detected,17 cases(70.83%)were ureaplasma parvum,and 7 cases(29.17%)were ureaplasma urealyticum.and no co-existence of ureaplasma parvum and ureaplasma urealyticum was detected.18 cases of positive specimens were also tested in drug susceptibility testing,of which ciprofloxacin and ofloxacin showed the lowest activity.Conclusion Compared with in vitro culture combined with sequencing,in vitro culture and RNA isothermal amplification showed better sensitivity and specificity,but from the point of view of clinical application,the combination of the two assays can provide more comprehensive guidance for the clinical diagnosis and treatment.
Objective To compare the detection of Ureaplasma determined by in vitro culture and RNA isothermal amplification.Methods From January to August 2016,a total of 103 urine specimens were collected from outpatients of Peking Union Medical College Hospital.The specimens were divided into three parts for the detection of Ureaplasma.The first part was determined by RNA isothermal amplification assay,the second part was tested by in vitro culture,and the third part was detected by in vitro culture and sequencing.In vitro culture combined with sequencing will be used as a reference method to evaluate the detection ability of Ureaplasma tested by in vitro culture and RNA thermostatic amplification.Results Ureaplasma was detected in24 specimens(23.30%)based on in vitro culture and sequencing.Among them,14 specimens(17.50%)were male and 10 specimens(43.47%)were female,the difference was statistically significant(P=0.022).From the age group,the highest Ureaplasma detection rate was found in 40-<50 age group(5/12,41.67%).Compared with the reference method,the sensitivity and specificity of in vitro culture were 75%(18/24)and100.00%(79/79),and the sensitivity and specificity of RNA isothermal amplification were 95.83%(23/24)and 96.20%(76/79),respectively,the difference was not statistically significant(P>0.05).Of the 24 samples with positive ureaplasma detected,17 cases(70.83%)were ureaplasma parvum,and 7 cases(29.17%)were ureaplasma urealyticum.and no co-existence of ureaplasma parvum and ureaplasma urealyticum was detected.18 cases of positive specimens were also tested in drug susceptibility testing,of which ciprofloxacin and ofloxacin showed the lowest activity.Conclusion Compared with in vitro culture combined with sequencing,in vitro culture and RNA isothermal amplification showed better sensitivity and specificity,but from the point of view of clinical application,the combination of the two assays can provide more comprehensive guidance for the clinical diagnosis and treatment.
引文
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[13]LEE M Y,KIM M H,LEE W I,et al.Prevalence and antibiotic susceptibility of mycoplasma hominis and ureaplasma urealyticum in pregnant women[J].Yonsei Med J,2016,57(5):1271-1275.
[14]XIAO L,GLASS J I,PARALANOV V,et al.Detection and characterization of human ureaplasma species and serovars by Real-Time PCR[J].J Clin Microbiol,2010,48(8):2715-2723.
[15]王辉,郑义,王玮蓁,等.孕妇尿中微小脲原体和解脲脲原体的分种和分型[J].同济医科大学学报,2001,30(6):580-582.
[16]PICCINELLI G,GARGIULO F,BISCARO V A,et al.A-nalysis of mutations in DNA gyrase and topoisomeraseⅣof Ureaplasma urealyticum and Ureaplasma parvum serovars resistant to fluoroquinolones[J].Infect Gene Evol,2017,47(1):64-67.
[17]SONG J,QIAO Y,KONG Y,et al.Frequent topoisomeraseⅣmutations associated with fluoroquinolone resistance in Ureaplasma species[J].J Med Microbiol,2015,64(11):1315-1320.
[18]KAMIYA Y,SHIMADA Y,ITO S,et al.Analysis of the quinolone-resistance determining region of the gyrA gene and the analogous region of the parC gene in Ureaplasma parvum and Ureaplasma urealyticum detected in first-void urine of men with non-gonococcal urethritis[J].J Anti Chem,2013,68(2):480-482.
[19]SCHNEIDER S C,TINGUELY R,DROZ S,et al.Antibiotic susceptibility and sequence type distribution of ureaplasma species isolated from genital samples in Switzerland[J].Anti Agents Chem,2015,59(10):6026-6031.
[2]JALIL N,DOBLE A,GILCHRIST C,et al.Infection of the epididymis by Ureaplasma urealyticum[J].Genitourin Med,1988,64(6):367-368.
[3]GRENABO L,HEDELIN H,PETTERSSON S.Urinary infection stones caused by Ureaplasma urealyticum:a review[J].Scand J Infect Dis Suppl,1988,53(1):46-49.
[4]BACZYNSKA A,SVENSTRUP H F,FEDDER J,et al.The use of enzyme-linked immunosorbent assay for detection of Mycoplasma hominis antibodies in infertile women serum samples[J].Hum Rep,2005,20(5):1277-1285.
[5]CHAIM W,HOROWITZ S,DAVID J B,et al.Ureaplasma urealyticum in the development of postpartum endometritis[J].Eur J Obstet Gynecol Rep Biol,2003,109(2):145-148.
[6]WAITES K B,KATZ B,SCHELONKA R L.Mycoplasmas and ureaplasmas as neonatal pathogens[J].Clin Microbiol Rev,2005,18(4):757-789.
[7]HUANG C,LONG X Y,JING S,et al.Ureaplasma urealyticum and Mycoplasma hominis infections and semen quality in 19,098infertile men in China[J].World JUrol,2016,34(7):1039-1044.
[8]ZHANG L,ZHANG K P,LIANG C Z.Ureaplasma urealyticum in male genital tract:a hidden risk factor for male infertility[J].Andrologia,2016,48(10):1077-1079.
[9]LIU J J,WANG Q X,JI X F,et al.Prevalence of ureaplasma urealyticum,mycoplasma hominis,chlamydia trachomatis infections,and semen quality in infertile and fertile men in China[J].Urology,2014,83(4):795-799.
[10]LEE J S,KIM K T,LEE H S,et al.Concordance of ureaplasma urealyticum and mycoplasma hominis in infertile couples:impact on semen parameters[J].Urology,2013,81(6):1219-1224.
[11]ZENG X Y,XIN N,TONG X N,et al.Prevalence and antibiotic susceptibility of Ureaplasma urealyticum and Mycoplasma hominis in Xi′an,China[J].Eur J Clin Microbiol Infect Dis,2016,35(12):1941-1947.
[12]FERNANDEZ J,KARAU M J,CUNNINGHAM S A,et al.Antimicrobial susceptibility and clonality of clinical ureaplasma isolates in the United States[J].Anti Agents Chem,2016,60(8):4793-4798.
[13]LEE M Y,KIM M H,LEE W I,et al.Prevalence and antibiotic susceptibility of mycoplasma hominis and ureaplasma urealyticum in pregnant women[J].Yonsei Med J,2016,57(5):1271-1275.
[14]XIAO L,GLASS J I,PARALANOV V,et al.Detection and characterization of human ureaplasma species and serovars by Real-Time PCR[J].J Clin Microbiol,2010,48(8):2715-2723.
[15]王辉,郑义,王玮蓁,等.孕妇尿中微小脲原体和解脲脲原体的分种和分型[J].同济医科大学学报,2001,30(6):580-582.
[16]PICCINELLI G,GARGIULO F,BISCARO V A,et al.A-nalysis of mutations in DNA gyrase and topoisomeraseⅣof Ureaplasma urealyticum and Ureaplasma parvum serovars resistant to fluoroquinolones[J].Infect Gene Evol,2017,47(1):64-67.
[17]SONG J,QIAO Y,KONG Y,et al.Frequent topoisomeraseⅣmutations associated with fluoroquinolone resistance in Ureaplasma species[J].J Med Microbiol,2015,64(11):1315-1320.
[18]KAMIYA Y,SHIMADA Y,ITO S,et al.Analysis of the quinolone-resistance determining region of the gyrA gene and the analogous region of the parC gene in Ureaplasma parvum and Ureaplasma urealyticum detected in first-void urine of men with non-gonococcal urethritis[J].J Anti Chem,2013,68(2):480-482.
[19]SCHNEIDER S C,TINGUELY R,DROZ S,et al.Antibiotic susceptibility and sequence type distribution of ureaplasma species isolated from genital samples in Switzerland[J].Anti Agents Chem,2015,59(10):6026-6031.