Locked nucleic acid real-time polymerase chain reaction method identifying two polymorphisms of hepatitis B virus genotype C2 infections,rt269L and rt269I

BACKGROUND The presence of two distinct hepatitis B virus(HBV)Pol RT polymorphisms,rt269L and rt269I,could contribute to the unique clinical or virological phenotype of HBV genotype C2.Therefore,a simple and sensitive method capable of identifying both types in chronic hepatitis B(CHB)patients infected with genotype C2 should be developed.AIM To develop a novel simple and sensitive locked nucleic acid(LNA)-real timepolymerase chain reaction(RT-PCR)method capable of identifying two rt269 types in CHB genotype C2 patients.METHODS We designed proper primer and probe sets for LNA-RT-PCR for the separation of rt269 types.Using synthesized DNAs of the wild type and variant forms,melting temperature analysis,detection sensitivity,and endpoint genotyping for LNA-RT-PCR were performed.The developed LNA-RT-PCR method was applied to a total of 94 CHB patients of genotype C2 for the identification of two rt269 polymorphisms,and these results were compared with those obtained by a direct sequencing protocol.RESULTS The LNA-RT-PCR method could identify two rt269L and rt269I polymorphisms of three genotypes,two rt269L types[‘L1’(WT)and‘L2’]and one rt269I type(‘I’)in single(63 samples,72.4%)or mixed forms(24 samples,27.6%)in 87(92.6%sensitivity)of 94 samples from Korean CHB patients.When the results were compared with those obtained by the direct sequencing protocol,the LNA-RT-PCR method showed the same results in all but one of 87 positive detected samples(98.9%specificity).CONCLUSION The newly developed LNA-RT-PCR method could identify two rt269 polymorphisms,rt269L and rt269I,in CHB patients with genotype C2 infections.This method could be effectively used for the understanding of disease progression in genotype C2 endemic areas.

Rapid genotyping of human rotavirus using SYBR green real-time reverse transcription-polymerase chain reaction with melting curve analysis

AIM: To develop a real-time reverse transcriptionpolymerase chain reaction(RT-PCR) assay to genotype rotavirus(G and P) in Alberta from January 2012 to June 2013. METHODS: We developed and validated a different approach to perform rotavirus G and P genotyping using a two-step SYBR green RT-PCR(rt-g PCR) by selecting genotype-specific primers of published conventional RT nested PCR(cn RT-PCR) assay and optimizing the amplification conditions. c DNA was first synthesized from total RNA with Super Script? Ⅱ reverse transcriptase kit followed by amplication step using monoplex SYBR green real-time PCR. After the PCR reaction, melting curve analysis was used to determine specific genotype. Sixteen samples previously genotyped using cn RT-PCR were tested using the new assay and the genotyping results were compared as sensitivity analysis. Assay specificity was evaluated by testing other gastroenteritis viruses with the new assay. The amplicon size of each available genotype was determined by gelelectrophoresis and DNA sequences were obtained using Sanger-sequencing method. After validation and optimization, the new assay was used to genotype 122 pediatric clinical stool samples previously tested positive for rotavirus using electron microscopy between January2012 and June 2013.RESULTS: The new rt-g PCR assay was validated and optimized. The assay detected G1 to G4, G9, G12 and P[4] and P[8] that were available as positive controls in our laboratory. A single and clear peak of melting curve was generated for each of specific G and P genotypes with a Tm ranging from 80 ℃ to 82 ℃. The sensitivity of rt-g PCR was comparable to cn RT-PCR with 100% correlation of the 16 samples with known G and P genotypes. No cross reaction was found with other gastroenteritis viruses. Using the new rt-g PCR assay, genotypes were obtained for 121 of the 122 pediatric clinical samples tested positive for rotavirus: G1P[8](42.6%), G2P[4](4.9%), G3P[8](10.7%), G9P[8](10.7%), G9P[4](6.6%), G12P[8](23.0%), and unknown GP[8](0.8%). For the first time, G12 rotavirus strains were found in Alberta and G12 was the second most common genotype during the study period. Gel electrophoresis of all the genotypes showed expected amplicon size for each genotype. The sequence data of the two G12 samples along with other genotypes were blasted in NCBI BLAST or analyzed with Rota C Genotyping tool(http://rotac.regatools.be/). All genotyping results were confirmed to be correct.CONCLUSION: rt-g PCR is a useful tool for the genotyping and characterization of rotavirus. Monitoring of rotavirus genotypes is important for the identification of emerging strains and ongoing evaluation of rotavirus vaccination programs.

Evaluation of patients with dry eye disease for conjunctival Chlamydia trachomatis and Ureaplasma urealyticum

AIM：To determine the possibility of the development of dry eye disease（DED） as a result of persistent infection with Chlamydia trachomatis and Ureaplasma urealyticum in the conjunctiva of patients.METHODS： This study was conducted on 58 patients of age range 20-50 y,diagnosed with DED confirmed by Schirmer I test and tear breakup time.The non-dry eye control group included 27 subjects of the same age.Ocular specimens were collected as conjunctival scrapings and swabs divided into three groups： the first used for bacterial culture,the second and third taken to detect Chlamydia trachomatis and Ureaplasma urealyticum by direct fluorescent antibody（DFA） assay and polymerase chain reaction（PCR） method. RESULTS： Chlamydia trachomatis was detected in 65.5% and 76% of DED patients by DFA and PCR methods respectively.Ureaplasma urealyticum was found in 44.8% of DED infected patients using the PCR method.Both organisms were identified in only 37.9% of DED patients found to be infected.Control subjects had a 22%detection rate of Chlamydia trachomatis by DFA assay versus a 7% detection rate by PCR; while Ureaplasma urealyticum was detected in 3.7% of the controls by PCR method.The conjunctival culture revealed that gram positive microorganisms represented 75% of isolates with coagulase negative Staphylococci the most common（50%） followed by Staphylococcus aureus（20%）,whereas gram negative microorganisms occurred in 25% of cases,isolating Moraxella spp.as the most frequent organism. CONCLUSION： Our results tend to point out that Chlamydia trachomatis and Ureaplasma urealyticum were detected in a moderate percentage of patients with DED,and could be a fair possibility for its development.PCR is more reliable in detecting Chlamydia trachomatis than DFA technique.The presence of isolated conjunctival bacterial microflora can be of some potential value.

Study of the prevalence and association of ocular chlamydial conjunctivitis in women with genital infection by Chlamydia trachomatis, Mycoplasma genitalium and Candida albicans attending outpatient clinic

AIMTo determine the association between chlamydial conjunctivitis and genital infection by Chlamydia trachomatis, Mycoplasma genitalium and Candida albicans, in addition to the possible relationship between cultured bacterial pathogens and oculogenital chlamydial infection.METHODSThis study was performed on 100 (50 symptomatic and 50 asymptomatic) women attending the Gynecological and Obstetric outpatient clinic of Alzahra hospital, Alazhar University. Simultaneously a conjunctival swab was taken from these patients. Polymerase chain reaction (PCR) was done on DNA extracted from both vaginal and conjunctival swab samples. Culture for both vaginal and conjunctival swabs was also done.RESULTSCandida albicans was the predominant organism isolated by culture in 20% and 40% of conjunctival and vaginal swabs respectively. By the PCR method, ocular Chlamydia trachomatis was present in 60% of symptomatic women, while genital Chlamydia trachomatis infection was present in 30% of symptomatic women. The results of this method also indicated that 25/50 (50%) vaginal swabs were positive with PCR for Candida albicans versus 15/50 (30%) were PCR positive in conjunctival swab. Mycoplasma genitalium was present in only 10% of vaginal swabs. Concomitant oculogenital PCR positive results for Chlamydia trachomatis and Candida albicans were 30% and 28% respectively.CONCLUSIONOcular Chlamydia trachomatis was associated with genital Chlamydia trachomatis in a high percentage of women followed by Candida albicans. Cultured bacterial organisms do not play a role in enhancement of Chlamydia trachomatis infection.

Detection of <i>Chlamydia trachomatis</i>and <i>Neisseria gonorrhoeae</i>in Egyptian Women Suffering from Infertility

Chlamydial and gonococcal infections are recognized as two of the major causes of sexually transmissible human bacterial infection which may lead to infertility. In this cross sectional study, we aimed to determine the prevalence of Neisseria gonorrhoeae, Chlamydia trachomatis among Egyptian women using different microbiological methods. One hundred and fifty cervical swabs were collected, of which 100 were from infertile women. Culture and ELISA technique were used for screening of Neisseria gonorrhoeae and Chlamydia trachomatis individually. In addition, PCR was used for all examined samples. For C. trachomatis, 3 cases were positive for antigen detection by ELISA. Moreover, in obtained results of PCR, DNA was detected in 4 samples, and three of them from infertile group. So based on PCR results, the sensitivity and specificity of ELISA were 75% and 100% respectively. Furthermore, 3 samples were positive for gonococcal infections by PCR, and two of them were taken from infertile women. Positive results of two samples were verified by culture. The estimated sensitivity and specificity of culture method were 66.7% and 100% respectively. Results of this study indicate that PCR is a valuable method for detection of gonococcal and chlamydial infection and it is suitable for the confirmation of ELISA results for C. trachomatis diagnosis. Culture method is less sensitive than PCR for detection of N. gonorrhoeae. The prevalence of such infections is higher among infertile women.

Detection of Genitourinary Tract Chlamydia trachomatis Infection in Urine specimens by PCR Assay

Objective: To compare the sensitivity and specificity of the cervical/urethral swabs with voided urine specimens for the detection of genitourinary tract infection with Chlamydia trachomatis and determine whether urine specimens can replace the cervical/urethral swabs in detection of C. trachomatis. Methods: The matched cervical/urethral swabs and voided urine specimens were collected from 569 patients of STD clinics.Polymerase chain reaction (PCR) assay specific for C. trachomatis plasmid DNA and rapid antigen testing (Clear view assay) was used to detect C. trachomatis. Standard criteria that defined """"true"""" positive included: 1) positive PCR results both in cervical/urethral swab and voided urine specimen or 2) positive voided urine results both by PCR assay and clear view test or 3)positive results in both PCR assay of cervical/urethral swab and clear view test of voided urine. For statistical analysis, the chi-square test was used. Results: The prevalence of C. trachomatis in patients with symptoms was 12.1% (28/231) in women and 10.4%(10/96) in men, with no significant difference between them (x^2=0.21,P>0.05). The prevalence of C. trachomatis in patients with no symptoms was 11.0% (11/100) in women and 15.5% (22/142) in men, with a significant difference existing between them. (x^2=4.0, P<0.05). No significant difference (P>0.05) existed between PCR testing of swabs (sensitivity 87.3 %; specificity 99.2 %) and PCR testing of urine (sensitivity 88.7%; specificity 98.8%). As for clear view assay, sensitivity was 60.6% and specificity was 100%. Conclusions: PCR assay is superior to clear view in detecting C. trachomatis. Although both PCR testing of swabs and PCR testing of urine specimens both have high sensitivity and specificity, urine specimen testing is more cost-effective, practical and noninvasive. Thus urine specimens can take the place of the swabs in PCR testing for chlamydia.

北京朝阳区28923例女性人乳头瘤病毒感染基因型分析

目的分析北京朝阳区28923例人乳头瘤病毒(HPV)检测结果的感染情况、基因分型及在各年龄段的分布差异,为本地区的宫颈癌(CC)防治及疫苗推广提供参考依据。方法回顾性分析2019年1月至2021年12月在首都医科大学附属北京妇产医院/北京妇幼保健院妇科肿瘤科、计划生育科、妇保妇检科等科室首次就诊且自愿进行多重荧光PCR技术23种HPV基因分型检测的28923例女性患者的人乳头瘤病毒筛查检测结果。结果28923例患者中,检测出HPV感染者11076例,感染率为38.29%;所有HPV感染者中单一型别感染占比24.45%、两重感染占比8.56%、双重以上型别感染占比5.27%;患者年龄从低到高感染率趋势呈U型分布;单一型别感染中高危型HPV(HR-HPV)中最常见的基因型是HPV16型,其次是52、58、53型,单一型别感染中低危型HPV(LR-HPV)中最常见的基因型是HPV81型,其次是42、44、43型;28923例感染者中31~40岁年龄组患者人数最多为8958例(占比31%);检出率小于20岁年龄组最高为55.56%,其次为21~30岁年龄组为42.62%,接下来是大于60岁年龄组为41.88%。结论北京朝阳区总体HR-HPV中感染率最高型别依次是HPV16、52及58型,LR-HPV中感染率最高的型别依次是HPV42、81及44型;年龄上31~50岁组感染率更高,单一HPV感染率高于多重感染,应提高本地区HPV16、52、58型的筛查能力,引起足够重视,有针对性的推广CC疫苗的接种,以便有效降低本地区CC的发生。

Helicobacter pylori vacA genotypes and cagA status and their relationship to associated diseases

INTRODUCTIONHelicobacter pylori(H.pylori)is a major causativebacterium of chronic gastritis,peptic ulcer and mucosa-associated lymphoid tissue lymphoma in humans,andassociated with an increased risk of gastric cancer.

Relationship between genotype and phenotype of flagellin C in Salmonella

AIM: To discover the relationship between the genotype and antigen serotype of flagellin C among Salmonella strains.METHODS: Fragment of Salmonella flagellin C in plasmid pLS408 was cloned, sequenced and compared with the corresponding sequence in other strains. Salmonella strains including two typhi strains, one paratyphoid strain, one enteritidis and one typhimurium strain were isolated from outpatients. Genome DNA was purified respectively from these clinical isolstes, then the corresponding flagellin C fragment was amplified by polymerase chain reaction, and the amplification products were analyzed by agarose gel electrophoreeis.RESULTS: The cloned fragment includes 582 nucleotides encoding the variable region and partial conservative region of Salmonella flagellin C in plasmid pLS408. With comparison to the corresponding sequences reported previously, there is only a little difference from other strains with the same flagellar serotype in both nucleotide and amino acid level. Specific PCR products were amplified in Salmonella strains with flagellar eerotype H-1-d including S. Muenchen, typhi and typhimurium, but not in S.paratyphoid C or S. Enteritidis strains.CONCLUSION: In this experiment, the specificity of nucleotide sequence could be found in flagellin C central variable regions as it exists in flagellar serotypes in Salmonella. It may be helpful to developing a rapid,sensitive, accurate and PCR-based method to detect Salmonella strains with serotype H-1-d.

Prevalence of Helicobacter pylori virulence genotypes among children in Eastern Turkey

AIM:To identify the virulence genotypes of Helicobacter pylori(H.pylori)if present in children in Eastern Turkey and if those genotypes are mostly associated with severe clinical presentations.METHODS:A total of 49 H.pylori positive Turkish children(42 with antral nodularity and 7 with peptic ulcer)who underwent upper gastrointestinal endoscopy with abdominal symptoms during the period from March 2011 to September 2012 were enrolled in this study.Antral nodularity was diagnosed endoscopically by two of the authors.We determined for the presence of cagA,vacA,cagE,iceA and babA2 genotypes of H.pylori isolates in DNA obtained directly from frozen gastric biopsy samples by polymerase chain reaction test using specific primers.RESULTS:Of the 49 H.pylori isolates studied,61.2%,91.8%,22.4%,28.6%,57.1%and 40.8%were positive for the cagA,vacA s1,cagE,iceA1,iceA2 and babA2 genes,respectively.We showed that the most common vacA subtype was s1a(79.6%).However,the s2 gene was found less frequently with an isolation rate of 8.2%of the H.pylori isolates.The genotypes iceA2 and vacA s1m2 were the most frequently found types in children with antral nodularity.In addition,the genotypes iceA1,babA2 and vacA s1m1 were found in similar ratios in all the H.pylori isolates obtained from children with peptic ulcer.The genotypes vacA s2m1and s1c were not observed in any of isolates studied.CONCLUSION:This study showed that vacA s1m2,cagA and iceA2 were the most common genotypes,and no association between antral nodularity and genotypes was observed.

Research on Hepatitis B virus Genotypes and Subgenotypes among Bai Nationality in Dali, Yunnan Province

To investigate the distribution of hepatitis B virus (HBV) genotypes and subgenotypes among the Bai nationality in Dali, a total of 100 serum samples from patients with chronic HBV-infection were collected for the detection of HBV genotypes and subgenotypes by genotype-specific primers and restriction fragment length polymorphism (RLFP), respectively. Among the 100 samples, the proportions of genotype B, C and mixed genotype (B+C) were 41%, 25% and 34%, respectively. All the genotype B strains belonged to subgenotype Ba. In genotype C, 84% were Subgenotype Cs and 12% were subgenotype Ce. The distribution of genotypes B, C and B+C showed no significant difference between male and female patients (P=0.182) and among the age groups of patients (P=0.812). The rates of HBeAg/HBeAg positivity were no significantly different among genotypes B, genotype C and mixed genotype (B+C) (P=0.077/P=0.663). In Dali, genotypes B, B+C and C existed among Bai nationality with chronic HBV-infection, and genotype B was the major genotype. Subgenotypes Ba and Cs were the predominant strains in patients with HBV genotype B/C infection. The most prominent characteristic was the higher prevalent rate of mixed genotype (B+C) in patients.

Detection of Chlamydia pneumonia DNA in nasopharyngolaryngeal swab samples from patients with rhinitis and pharyngolaryngitis with polymerase chain reaction

Objective To assess the prevalence of Chlamydia pneumomia DNA in patients with otolaryngic disease Methods PCR assay was used to detect Chlamydia pneumonia specific Pst Ⅰ 474 fragment DNA in swabs from patients with acute or subacute pharyngolaryngitis or rhinitis and sinusitis C pneumonia specific antibodies in sera were also assayed with microimmuno fluoresence (MIF) Results About 28% (49/175) of the patients were PCR positive and 25 7%(45/175) were MIF antibodies positive The accordance rate of the two methods was 91 8% Conclusion It is suggested that the C pneumonia infection was common in this group of patients and the C pneumonia Pst Ⅰ474 specific PCR was sensitive and specific for detecting C pneumonia in pharyngolaryngitis or rhinitis and sinusitis

Follow up of infection of chacma baboons with inoculum containing a and non-a genotypes of hepatitis B virus

AIM: To determine whether one genotype (A or non-A genotypes of HBV) predominated over the other during the course of HBV infection.METHODS: Four baboons were inoculated with HBV. DNA was extracted from serum obtained at monthly intervals postinoculation for 52 weeks and HBV DNA was amplified using primers specific for the core region containing an insert characteristic of genotype A (nt 2 354-2 359, numbering from the EcoRI site). The amplicons were cloned into PCRScriptTM and a minimum of 15 clones per time point were sequenced in both directions.RESULTS: Both genotypes persisted for the entire followup period of 52 weeks. Genotype non-A predominated in two baboons and genotype A in one baboon. Neither genotype predominated in the fourth baboon, as shown at a 5 % level of testing.CONCLUSION: No conclusions concerning the dominance of either genotype or the natural progression or replication rates of HBV could be drawn because the pattern of the genotypes found may have been caused by sampling fluctuations at the time of DNA extraction and cloning as a result of the very low viral loads in the baboon sera.

2017—2019年福建省结核分枝杆菌分离株基因型特征及其耐药性分析

目的:了解福建省结核分枝杆菌(Mycobacterium tuberculosis,MTB)基因型的分布特征和流行情况,同时分析MTB基因型与其耐药的关系。方法:选取2017—2019年福建省结核病耐药监测点的477株MTB临床分离株作为研究对象。采用对硝基苯甲酸(PNB)/噻吩-2-羧酸肼(TCH)生长实验法进行菌种初步鉴定,并采用传统固体比例法对9种抗结核药物[异烟肼(isoniazid,INH)、利福平(rifampin,RFP)、链霉素(streptomycin,Sm)、乙胺丁醇(ethambutol,EMB)、卡那霉素(kanamycin,Km)、氧氟沙星(ofloxacin,Ofx)、卷曲霉素(capreomycin,Cm)、丙硫异烟胺(prothionamide,Pto)、对氨基水杨酸(para-aminosalicylic acid,PAS)]进行药物敏感性试验(简称“药敏试验”)。应用熔解曲线法间隔区寡核苷酸分型(McSpoligotyping)技术对菌株进行基因分型。结果:477株MTB菌株中北京基因型有245株,占51.4%;T家族有44株(含2株T,28株T1,7株T2,T3和T4各3株,1株T5),占9.2%;H家族有35株(含13株H,1株H1,21株H3),占7.3%;EAI家族有11株(含10株EAI2-Manila,1株EAI5),占2.3%;LAM3、Manu2和X1家族各有1株,分别占0.2%;所属家族未定义或不明确的有139株,占29.1%。聚类分析显示,各基因型家族主要流行型为北京家族的SIT1(44.9%,214/477)、T家族的SIT53(2.5%,12/477)、H家族的SIT742(1.9%,9/477)、EAI家族的SIT19(1.5%,7/477)。各家族MTB菌株与INH的耐药率之间的差异有统计学意义(χ^(2)=10.311,P=0.036),其中,EAI+LAM+Manu+X家族菌株对INH的耐药率最高[28.6%(4/14)];对RFP的耐药率之间的差异有统计学意义(χ^(2)=14.366,P=0.006),其中,EAI+LAM+Manu+X家族菌株对RFP的耐药率最高[21.4%(3/14)];对Ofx的耐药率之间的差异有统计学意义(χ^(2)=23.643,P=0.000),其中,H家族菌株对Ofx的耐药率最高[17.1%(6/35)];对PAS耐药率之间的差异有统计学意义(χ^(2)=9.550,P=0.049),其中,未定义家族菌株对PAS的耐药率最高[4.3%(6/139)]。结论:福建省MTB流行基因型以北京基因型为主,同时应重视未定义家族菌株的流行并加强对T家族、H家族和EAI等家族菌株的监测。菌株对INH、RFP、Ofx和PAS的耐药性与各基因家族相关。

Relationship between Chlamydia pneumoniae infection and occurrence of bronchial asthma

Objective: To study the relationship between Chlamydia pneumoniae (C. pneumoniae) infection and asthma exacerbation. Methods: A prospective study of C. pneumoniae infection was conducted in 75 patients with asthma and 63 patients with respiratory tract infection, and 100 blood donors served as controls. The presence of infection was convinced by the polymerase chain reaction and direct immunofluorescence assay for C. pneumoniae DNA from throat swab specimens and micro-immunofluorescence testing for C. pneu-moniae-specific IgG, IgM and IgA antibodies. Results: Prevalence of specific IgG in asthma patients (81. 3%) was higher than that of the blood donors (68. 0%, P<0. 05) and was not significantly different from respiratory tract infection patients (68. 0%, P>0. 05). The acute C. pneumoniae infection rate of symptomatic asthma patients (59. 4%) was markedly higher than that of respiratory tract infection patients (34. 9% , P<0. 05). The average titer of C. pneumoniae IgG instead of IgA in asthma patients (48.

Relationship between Glucocorticoid-induced Osteoporosis and Vitamine D Receptor Genotypes

By means of polym erase chain reaction- restriction fragment length polymorphism (PCR- RFL P) assay,the association between vitamine D receptor (VDR) genotypes and bone min- eral density (BMD) in the patients receiving long- term glucocorticoid therapy was studied.The clinical data and blood of71patients with rheumatosis who received long- term glucocorticoid ther- apy were collected.BMD was m easured by dual- energy X- ray absorptimometry.VDR gene frag- ment(about185 bp) was amplified by PCR from the extracted genomic DNA,then digested with restriction endonuclease Bsm I.The genotypes were evaluated based on the fragment length fol- lowing endonuclease digestion and the association between genotypes and BMD or Z- score values was analyzed.Among the 71cases,the detected genotypes were Bb and bb with the distribution frequency being 11.3% and 88.7% respectively.The distribution frequency of the alleles was in agreement with the Hardy- Weinberg equilibrium.There was no significant difference between the two genotypes in age,gender,body m ass index(BMI) ,disease duration,disease types,time of glucocorticoid administration and cumulative dosage(P>0 .0 5 ) .Osteoporosis rate of the patients with Bb or bb genotype was37.5 % and33.3% respectively,with the difference being notsignif- icant (χ2 =0 .0 5 ,P=0 .8) .The BMD and Z- score values at lumbar spine and femur in two geno- types were not similar,but the difference had no significant (P>0 .0 5 ) .The distribution frequen- cy of bb type of VDR genotypes in Han populations of China was m ore prevalent,followed by Bb and bb types in turn.In the patients receiving long- term glucocorticoid therapy,there was no sig- nificant difference in BMD between Bb and bb genotypes.The data suggest that the VDR geno- types may not be m eans of identifying patients at greater risk of glucocorticoid- induced osteoporo- sis,which await to be further confirmed by a large sample size.

荧光定量PCR与全自动生物芯片法检测人乳头瘤病毒的一致性研究

背景 人乳头瘤病毒(human papillomaviruses,HPV)分型检测是宫颈癌筛查的重要手段之一,但技术人员短缺及实验室硬件设施不足对其应用造成了不利影响。目的 以荧光定量聚合酶链式反应(PCR)为参考,考察全自动生物芯片法在HPV分型检测中的诊断性能。方法 针对常规进行荧光定量PCR检测的临床样本,每一种型别均采用抽签法进行简单随机抽样,选中的样本使用全自动生物芯片法检测,采用Kappa检验比较两种方法的检测结果是否存在差异。针对不一致结果,通过E6/E7区的扩增产物测序进行确认。结果 从4 589例样本中选择124例,其中HPV阴性50例,HPV阳性74例,按照阳性型别计算共99个测试。按照样本阴性、阳性计算,两种方法的结果完全一致,Kappa值为1.000(P<0.000 1)。按照样本型别计算,全自动生物芯片法的敏感度、特异度、阳性预测值、阴性预测值和总体符合率分别为98.0%、92.6%、96.0%、96.2%和96.1%,Kappa值为0.913(P<0.000 1),提示两种方法的结果一致性良好。两种方法不一致的结果集中在6例阳性样本之中,39型和59型各1例,52型和68型各2例,在重复验证及测序分析中均得到了确认。此外,全自动生物芯片法还发现了18例不被荧光定量PCR方法涵盖的型别,能够为临床治疗提供更丰富的信息。结论 全自动生物芯片法与荧光定量PCR法在HPV的分型检测中具有很好的一致性,对于技术人员短缺及不具备标准PCR实验室的医疗机构,全自动生物芯片法具有良好的应用前景。