Laboratory testing techniques for monkeypox virus / 中华皮肤科杂志

Monkeypox is a zoonotic viral disease caused by monkeypox virus infection. Monkeypox has become a public health emergency of international concern, since it first spread widely in many regions outside Africa in 2022. Accurate and effective detection methods are particularly important for the diagnosis and screening of monkeypox virus infection. This review summarizes laboratory testing techniques for monkeypox virus in recent years, and compares principles and detection performance of microscopy, culture, nucleic acid testing and immunological methods.

Strengthening comprehensive surveillance of and public health response to monkeypox in China / 中华皮肤科杂志

Monkeypox is a zoonotic disease caused by monkeypox virus, and human cases infected with the virus have been reported in more than 100 countries. To respond to the potential of case importation and consequent spread of the infection in the country, it is urgent for China to strengthen its comprehensive surveillance efforts consisting of case detection through country-entering check, symptom screening, and investigation among priority populations, and to implement comprehensive strategies to control the source of infection, interrupt the transmission and protect the people at risk.

Ceftriaxone-resistant Neisseria gonorrhoeae strain FC428: prevalence, resistance mechanisms and control strategies / 中华皮肤科杂志

Antimicrobial resistance of Neisseria gonorrhoeae has become a big challenge in the control and prevention of sexually transmitted diseases. Recently, a ceftriaxone-resistant Neisseria gonorrhoeae strain FC428 has spread across the world including China, which has worsened the antimicrobial resistance problem. This strain is highly resistant to ceftriaxone due to a novel mosaic penA gene. In order to better understand the characteristics of FC428 and control its further spread, this review summarizes its origin, spread, main molecular characteristics, resistance mechanisms, detection methods, and strategies for clinical treatment and antimicrobial resistance surveillance.

Evaluation of prototype IgA rapid test on diagnosis of early active syphilis / 中华皮肤科杂志

Objective To evaluate the application of Burnet Institute-made prototype IgA rapid test,a kind of point-of-care (POC) testing,in the diagnosis of early syphilis.Methods Totally,455 stored serum samples in the Reference Laboratory of Sexually Transmitted Disease,the Institute of Dermatology were used to evaluate the application of the prototype IgA rapid test (IgA-POC) in the diagnosis of early syphilis.According to resluts of Treponema pallidum hemagglutination assay (TPHA),rapid plasma reagin card test (RPR),and enzyme-linked immunosorbent assay for IgM antibodies (IgM-ELISA),these stored samples were divided into 3 groups:uninfected group,previously infected group and early active syphilis group.IgA-POC test was performed in the 3 groups to evaluate its diagnostic performance for active syphilis,and researchers were blind to the group information.Results The prototype IgA-POC test had a sensitivity of 92.6％ (147/163) for the early active syphilis group,a specificity of 72.22％ (104/144) for the previously infected group,and a specificity of 97.97％ (145/148) for the uninfected group.The total specificity of the prototype IgA-POC test was 85.27％,which met the minimum requirement of WHO for the POC test.The prototype IgA-POC test showed a significantly higher sensitivity for the diagnosis of early active syphilis compared with the IgM-ELISA (59.51％,Z =6.88,P ＜ 0.05),but a significantly lower specificity for the diagnosis of previous syphilis infection compared with the IgM-ELISA (98.61％,Z =6.18,P ＜ 0.05).Moreover,no significant difference in the specificity for the diagnosis of non-infection was observed between the prototype IgA-POC test and IgM-ELISA (Z =1.16,P =0.25).Conclusion The prototype IgA-POC test has better capacity for the diagnosis of early active syphilis compared with the IgM-ELISA,so it can be applied to the screening of early active syphilis.

In vitro combined effect of ceftriaxone and azithromycin against Neisseria gonorrhoeae / 中华皮肤科杂志

Objective To investigate in vitro combined effect of ceftriaxone and azithromycin against clinical isolates of Neisseria gonorrhoeae(NG). Methods A total of 25 NG clinical isolates were collected from the STD clinic of Dalian Dermatosis Hospital in 2012. Epsilometer test(Etest)method was used to determine the minimum inhibitory concentrations(MICs)of ceftriaxone and azithromycin against NG isolates. Fractional inhibitory concentration index (FICI) was calculated to evaluate the in vitro combined effect of ceftriaxone and azithromycin against NG isolates. Results The mean MICs of ceftriaxone and azithromycin were 0.032 mg/L (range, 0.008- 0.064 mg/L) and 0.834 mg/L (range, 0.064-4.000 mg/L), respectively. The FICI ranged from 0.724 to 2.696, and ceftriaxone and azithromycin showed an additive effect against the above NG isolates. Conclusion Ceftriaxone and azithromycin show an additive effect against NG in vitro, but further studies with large sample size are needed to confirm their effects.

High-resolution multilocus sequence typing of urogenital Chlamydia trachomatis among STD clinic outpatients in Guangxi Zhuang Autonomous Region / 中华皮肤科杂志

Objective To investigate the type distribution of urogenital Chlamydia trachomatis(Ct)among STD clinic outpatients in Guangxi Zhuang Autonomous Region, and to estimate the prevalence of Ct infection among the patients during posttreatment follow?up. Methods Urethral and cervical swabs were collected from male and female outpatients with confirmed urogenital Ct infection, respectively, in Institute of Dermatology of Guangxi Zhuang Autonomous Region. The patients with positive results in preliminary screening tests were followed up after treatment, and specimens were collected at follow?up visits. General and clinical information was also obtained from these patients. DNA was extracted from these samples by using the QIAxtractor instrument. Nested PCR was performed to amplify the major outer membrane protein A(ompA)gene for ompA typing, and to amplify CT046(hctB), CT058, CT144, CT172 and CT682 (pbpB) genes for high?resolution multilocus sequence typing (hr?MLST). Then, PCR products were sequenced, and ompA and MLST types of Ct were determined by sequence alignment and MLST analysis, respectively. The obtained MLST sequence types (STs) were compared with those from an Italian population by using the BioNumerics7 software, and a minimum spanning tree(MST)was generated. Results Totally, 44 and 6 Ct?positive specimens were collected at first visits and follow?up visits respectively. Among the 50 specimens, 42 underwent successful ompA typing and hr?MLST, and 7 ompA genotypes and 15 hr?MLST STs were identified, including 3 first reported STs. The distribution of STs of Ct isolates from Guangxi Zhuang Autonomous Region was significantly different from that from the Italian population. Among the 6 followed patients with posttreatment Ct infection, 3 were confirmed to be reinfected with Ct, and the other 3 failed to be diagnosed because of unsuccessful genotyping. Conclusion The genotypes of Ct strains isolated from STD clinic outpatients in Guangxi Autonomous Region were characteristic, and Ct reinfection occurred in some patients during follow?up.

Pathogenicity of Treponema pallidum inoculated through different routes in rabbits：a preliminary study / 中华皮肤科杂志

Objective To observe clinical manifestations and immune responses in rabbit models of syphilis established by inoculation with Treponema pallidum through different routes. Methods A total of 20 rabbits were randomly and equally divided into two groups: testis-inoculated group injected with Treponema pallidum (Tp) suspensions into the testes, back-inoculated group intracutaneously injected with Tp suspensions at six sites on the shaved back. All the rabbits received a single injection with the total injection amount of Tp (Nichols strains)being 2 × 107 in both groups. Clinical symptoms and immune responses were evaluated in both groups every other day. Statistical analysis was carried out by a two-sample t-test and rank sum test with the SPSS 20.0 software. Results On day 8 after the inoculation, the testes of rabbits in the testis-inoculated group started to swell with induration, and the swelling was most severe during days 13 - 16. Afterwards, the testes gradually diminished, and returned to normal in size without swelling or hardening on day 28. No skin lesions occurred in the other sites in the testis-inoculated group within 2 months after the inoculation. Erythematous swelling occurred at inoculation sites on day 7 after inoculation in the back-inoculated group, which was most obvious between days 15 and 45. Moreover, cartilage-like indurated ulcers were observed at 12 inoculation sites in 3 rabbits in the back-inoculated group, and dark-field examination of the ulcers showed the presence of Tp. There was a significant difference in the time required for the severity of lesions to peak between the testis-inoculated group and back-inoculated group (14.50 ± 1.08 days vs. 29.00 ± 10.30 days, t=5.02, P<0.05). Rank sum test showed significant differences in the distribution of highest RPR titers between the two groups(P<0.05), and RPR titers were higher in the testis-inoculated group than in the back-inoculated group. Conclusions The injection of Tp suspensions at different sites can cause different clinical symptoms and immune responses in rabbits. RPR turned positive earlier with a higher titer in the testis-inoculated group compared with the back-inoculated group. The clinical manifestations in the back-inoculated group were similar to those of chancre in primary syphilis in human.

Effects of medroxyprogesterone acetate and compound norethisterone enanthate on the susceptibility of BABL/c mice to lower reproductive tract infection with Chlamydia trachomatis / 中华皮肤科杂志

Objective To compare the effects of medroxyprogesterone acetate (MPA) and compound norethisterone enanthate (CNE) on the susceptibility of BABL/c mice to lower reproductive tract infection with chlamydia trachomatis (Ct). Methods A total of 60 BALB/c mice were randomly and equally divided into 6 groups:MPA-pretreated control group and CNE-pretreated control group inoculated with MyCoy cell suspensions in the vagina on the 5th day after single treatment with MPA and CNE respectively, blank control group receiving no treatment, MPA-pretreated infected group and CNE-pretreated infected group inoculated with 1 × 107 inclusion-forming units(IFU)of Ct serovar E in the vagina on the 5th day after single treatment with MPA and CNE respectively, control infected group inoculated with the same quantity of IFU of Ct serovar E in the vagina but receiving no pretreatment. On day 4, 7 and 14 after inoculation, vaginal irrigation fluid was obtained from all the mice for cell culture of Ct. Three mice were randomly selected from each of these groups at the above three time points and sacrificed, and vaginal and uterine tissue specimens were obtained for hematoxylin-eosin(HE)staining and microscopic examination. Chi-square test and Fisher's exact test were conducted to compare infection rate among different groups. Results No growth of Ct was observed in the three control groups at the above time points. The culture-positive rate of Ct was 1/10 on day 4 but 0 on day 7 and 14 in both the CNE-pretreated infected group and control infected group, 7/10 on day 4, 2/7 on day 7 but 0 on day 14 in the MPA-pretreated infected group. Fisher's exact test revealed that the culture-positive rate of Ct was significantly higher in the MPA-pretreated infected group than in the control infected group and CNE-pretreated infected group on day 4 (both P =0.03), but similar among the three infected groups on day 7 (P = 0.23). Both the MPA-pretreated control group and infected group showed an increase in endovaginal mucus, thinning of vaginal stratified squamous epithelium, mucification of vaginal epithelium, presence of secretions in vaginal lumen and submucosal infiltration of a few inflammatory cells on day 4, 7 and 14, as well as appearance of pathological changes (including the presence of large quantities of purulent secretions in lumen, mild tissue edema and submucosal infiltration of a few inflammatory cells) in the vagina on day 4. Vaginal tissues were normal in both the CNE-pretreated infected group and control group at the above three time points, but mild tissue edema, lumen expansion, secretion retention and infiltration of scattered inflammatory cells were observed in the uterus on day 4 after inoculation. Conclusions MPA can arrest the estrous cycle of mice at diestrus with the mucification of vaginal epithelium, which may increase the susceptibility to Ct vaginal infection in mice. In contrast, CNE has no obvious effect on the estrous cycle and susceptibility to Ct vaginal infection despite of the appearance of pathological changes in the uterus.

Penicillinase-producing Neisseria gonorrhoeae and its blaTEM-135 gene variants at several gonococcal antimicrobial surveillance sites in China：an epidemiological study / 中华皮肤科杂志

Objective To determine the prevalence of penicillinase-producing Neisseria gonorrhoeae(PPNG) and the distribution of blaTEM-135 gene variants in PPNG at several gonococcal antimicrobial surveillance sites in China, to compare N. gonorrhoeae multi-antigen sequence typing(NG-MAST)types of PPNG and its blaTEM-135 gene variants, and to assess the difference and association in NG-MAST types of blaTEM-135 gene variants among different regions. Methods A total of 572 N. gonorrhoeae isolates were collected at 6 gonococcal antimicrobial surveillance sites from Jiangsu, Shanghai, Zhejiang, Tianjin, Guangdong and Guangxi in 2012. After isolation, purification, and identification, cefalotin paper discs were used for detection of PPNG. DNA was extracted by QIAxtractor DX kits after cultivation of the PPNG strains. Then, mismatch amplification mutation assay (MAMA) PCR was performed to identify blaTEM-135 variants, and NG-MAST analysis to determine N. gonorrhoeae genotypes. Results Among the 572 N. gonorrhoeae strains, 38.1%(218/572) were identified as PPNG, and of the PPNG strains, 52.3% (114/218) were blaTEM-135 variants. The detection rate of PPNG at these surveillance sites from high to low was as follows: 51.7% (45/87, Zhejiang), 45.6%(36/79, Shanghai), 38.0% (78/205, Guangdong), 37.5% (12/32, Guangxi), 31.2% (24/77, Jiangsu) and 25.0%(23/92, Tianjin), and that of blaTEM-135 variants was as follows: 68.9%(31/45, Zhejiang), 58.3%(14/24, Jiangsu), 50.0%(39/78, Guangdong), 47.2%(17/36, Shanghai), 39.1%(9/23, Tianjin)and 33.3%(4/12, Guangxi). NG-MAST analysis showed that the ST2318, ST1768, ST1866, ST1053 and ST8726 types predominated among these bla TEM-135 variants, and a strong correlation was found between blaTEM-135 variants and some NG-MAST types, such as ST1768, ST1053 and ST8726 types. The distribution of NG-MAST types was significantly different between the surveillance site in Tianjin (in the Northern part of China) and the other sites (in the Southern part of China), but highly similar among the surveillance sites in Jiangsu, Zhejiang and Shanghai regions. Conclusions There is a high prevalence of PPNG and its blaTEM-135 variants at several gonococcal antimicrobial surveillance sites in China, with significant differences in NG-MAST genotype distribution of PPNG and its blaTEM-135 variants among different regions.

Ceftriaxone susceptibility testing and multi-antigen sequence typing of Neisseria gonorrhoeae strains isolated in 2007 and 2012 from Nanjing, China / 中华皮肤科杂志

Objective To test the ceftriaxone susceptibility of Neisseria gonorrhoeae (NG) isolates from Nanjing city,and to assess their genotypes by using the NG multi-antigen sequence typing (NG-MAST) method.Methods A total of 204 NG strains isolated in 2007 and 81 in 2012 from Nanjing city were included in this study.The minimum inhibitory concentration (MIC) of ceftriaxone was determined for these strains using an agar dilution method.DNA was extracted by the Qiagen commercial kit from these strains followed by NG-MAST.Results All the isolates were susceptible to ceftriaxone (MIC,≤ 0.25 μg/ml).The MIC of ceftriaxone was ≥ 0.06 μg/ml for 63.2％ of all the NG strains,70.6％ of those isolated in 2007 and 44.4％ of those in 2012,and ≥ 0.125 μg/ml for 31.6 ％ of all the NG strains,39.7％ of those isolated in 2007,11.1％ of those in 2012.Totally,166 genotypes were identified among the 285 isolates,of which,73 had been reported,and 93 were previously unreported.The most prevalent genotype was ST568 (n =13) in NG strains isolated in 2007,followed by ST270 (n =9),ST421 (n =7),ST2288 (n =5),ST1731 (n =4),ST1766 (n =4),ST1866 (n =4),ST1870 (n =4),while ST2318 (n =5),ST1053 (n =4),ST5990 (n =4),ST8726 (n =4) were the common genotypes in 2012.Those isolates with identical or similar genotypes tended to display similar MICs for ceftriaxone.Conclusions The prevalent genotypes of NG are markedly different between 2007 and 2012 in Nanjing region,and there is a strong association between the genotypes and ceftriaxone susceptibility of NG.NG-MAST results may serve as a genetic marker in the surveillance of antibiotic susceptibility in NG.

Pathogenicity of Ureaplasma urealyticum serotype 1 and 8 in the genital tract of female BALB/c mice / 中华皮肤科杂志

Objective To compare the pathogenicity between Ureaplasma urealyticum serotype 1 (Up1)and 8 (Uu8) in the genital tract of BALB/c mice.Methods A total of 48 BALB/c mice were randomly and equally divided into four groups:blank control group receiving no treatment,estradiol group pretreated with intramuscular injection of estradiol followed by intravaginal inoculation with sterial liquid culture media,Up1 and Uu8 groups pretreated with intramuscular injection of estradiol followed by intravaginal inoculation with suspensions of Up1 and Uu8 respectively.Three mice were randomly selected from each group to be sacrificed after the collection of vaginal lavage fluid on day 3,7,14 and 21 after the inoculation.Vaginal and uterine tissue specimens were obtained from these sacrificed mice and underwent hematoxylin and eosin (HE) staining.Vaginal lavage fluid samples were subjected to culture of Uu and measurement of tumor necrosis factor-α (TNF-α).Results No evidences were observed for Uu growth in either the blank control group or estradiol group at any of the time points after the inoculation,with the average level of TNF-α in vaginal lavage fluid being (4.17 ± 0.85) pg/ml at these time points in both groups.Uu grew in all the vaginal lavage fluid samples from the Up1 and Uu8 groups at the four time points,with the color change unit (CCU) value decreasing with time.The level of TNF-α in vaginal lavage fluid peaked on day 14 after the inoculation in the Up 1 ((14.93 ± 1.11) pg/ml) and Uu8 ((27.04 ± 24.26) pg/ml) groups.Both Up1 and Uu8 infection caused acute and chronic inflammatory responses in the mice,which were mainly located in the uterus,and Up1 might cause intrauterine adhesion.Conclusions At the same inoculation concentration,no significant difference is found in the pathogenicity between Up1 and Uu8,both of which appear to mainly cause cervicitis.Upl might be partially responsible for intrauterine adhesion in mice.

Effects of education level of men who have sex with men on their high risk sexual behaviors and the infection of HIV and syphilis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effects of educational background of men who have sex with men (MSM) on their high risk sexual behaviors and the HIV/STI infection rates.</p><p><b>METHODS</b>During July to November of 2009 and March to October of 2010, snowball and convenience sampling methods were adopted to recruit MSM from MSM venues and via the internet in Changzhou city of Jiangsu province, and finally 659 MSM were conducted a questionnaire survey and serological testing. According to the educational background of MSM, they were divided into 3 groups, that is, junior high school group (206 cases), high school group (254 cases), and university group (199 cases). The questionnaire mainly includes information on social demography, sexual behaviors, condom use, etc. Blood samples were collected for HIV and syphilis spirochete detection, and urine samples were also collected in 291 MSM who were recruited during July to November of 2009 for neisseria gonorrhoeae and chlamydia trachomatis nucleic acid detection. χ(2) test and other statistical analysis methods were used to compare the characteristics of sexual behaviors and HIV/STI infections in 3 groups.</p><p><b>RESULTS</b>Of the 659 valid questionnaires returned, junior high school group, high school group, and university group accounted for 31.3% (206 cases), 38.5% (254 cases) and 30.2% (199 cases). Places where MSM of different education levels most often to seek sexual partners, were significantly different. Junior high school group and high school group mostly went to bath house/sauna club (56.3%, 116 cases) and bar (34.8%, 88 cases) for partners, respectively, while the university group sought partners mainly through the internet (41.1%, 81 cases) (χ(2) = 99.35, P < 0.05). 53.4% (109/204) of the junior high school group had anal sex with men in the last 6 months, which was higher than that of high school group (67.7%, 172/254) (χ(2) = 9.74, P < 0.05) and university group (72.7%, 144/198) (χ(2) = 16.04, P < 0.05) . A total of 54.4% (111/204) of the junior high school group had sex with women in the last 6 months, which was higher than that of university group (38.6%, 76/197) (χ(2) = 10.10, P < 0.05) , but was not statistically significantly different from that of high school group (46.9%, 119/254) (χ(2) = 2.59, P = 0.11) . The rates of condom use with men at the last anal sex in junior high school group (73.4%, 80/109) , high school group (78.0%, 131/168) and university group (73.9%, 105/142) were similar. The rates of condom use with women in the last intercourse in junior high school group, high school group and university group were 51.8% (57/110), 54.6% (65/119) and 61.8% (47/76), respectively(χ(2) = 1.88, P = 0.39) . In junior high school group, high school group and university group, the infection rates of HIV were 9.2% (19/206), 10.6% (27/254) and 5.6% (11/197) (χ(2) = 3.68, P = 0.16), the positive rates of neisseria gonorrhoeae were 3.8% (3/79), 3.4% (4/117) and 0.0% (0/95) (χ(2) = 3.85, P = 0.14), the positive rates of chlamydia trachomatis were 5.1% (4/79), 9.4% (11/117) and 4.2% (4/95) (χ(2) = 2.70, P = 0.26). The infection rate of syphilis in junior high school group was 19.9% (41/206), which was higher than high school group (12.2%, 31/254) (χ(2) = 5.11, P < 0.05) and university group (10.2%, 20/197) (χ(2) = 7.45, P < 0.05 ).</p><p><b>CONCLUSION</b>There was no obvious correlation between education level and high risk sexual behaviors;MSM with lower education level were at higher risk of infection of syphilis.</p>

Nationwide external quality assessment of serologic tests for syphilis during 2006-2008 / 中华皮肤科杂志

Objective To make a nationwide external quality assessment of serologic tests for syphilis in China,in hope to increase the quality of syphilis serology in laboratories at different levels.Methods From 2006 to 2008,a nationwide external quality assessment scheme was conducted for serologic tests for syphilis in laboratories of some medical and healthcare facilities each year by the Reference Laboratory,National Center for Sexually Transmitted Disease Control,China Center for Disease Control and Prevention.Five quality control samples and corresponding questionnaires were sent to the participating laboratories.Tests were conducted and test results were reported within stipulated time.Subsequently,the test results were statistically analyzed by the Reference Laboratory,and the final results were fed back to all of these participants.Results From 2006 to 2008,the number of participating provinces increased from 17 to 31,and the number of participating laboratories from 23 to 145.Laboratories achieving a full score amounted to 79.9％,36.8％ and 57.6％,and those gaining a score of 80 or greater amounted to 95.7％,88.2％ and 89.7％,respectively,in 2006,2007 and 2008.Conclusion The external quality assessment scheme has enhanced the capacity of participating laboratories for syphilis serology to a certain extent from 2006 to 2008.

Prevalence of urogenital infection with and genotype distribution of Chlamydia trachomatis among female sex workers from different entertainment venues in Wuzhou and Hezhou cities of Guangxi Zhuang Autonomous Region / 中华皮肤科杂志

Objective To assess the prevalence of urogenital infection with and genotype distribution of C.trachomatis among female sex workers (FSWs) from different entertainment venues in Wuzhou and Hezhou cities of Guangxi Zhuang Autonomous Region.Methods A total of 810 FSWs were recruited to this study by convenience sampling from entertainment venues in Wuzhou and Hezhou cities of Guangxi Zhuang Autonomous Region from July 2009 to September 2010.Based on the venues where they solicited clients,the FSWs were classified into three tiers,i.e.,high-tier,middle-tier and low-tier.Cervical swabs were collected from all of these subjects followed by detection of C.trachomatis with the Amplicor PCR test kit.Then,DNA was extracted from C.trachomatis-positive specimens and subjected to nested PCR assay targeting the ompA gene followed by bidirectional sequencing.The genotype of C.trachomatis was determined according to the sequence of ompA gene.Chi-square test was conducted to compare the urogenital infection rate and genotype distribution of C.trachomatis between different tiers of FSWs.Results Among the 805 FSWs,the prevalence rate of urogenital C.trachomatis infection was 20.0％ (161/805).Chi-square test showed that the prevalence rate of urogenital C.trachomatis infection was significantly lower in high-and middle-tier FSWs than in low-tier FSWs (x2 =3.97,5.95,respectively,both P ＜ 0.05).Nine genotypes of C.trachomatis were identified in these FSWs,with serotype F as the most prevalent genotype (39/154,25.3％).Low-tier FSWs showed a higher frequency of genotype E (x2 =5.02,P ＜ 0.05) but a lower frequency of genotype K (Fisher's Exact test,P =0.048) compared with middle-tier FSWs.Conclusions Low-tier FSWs show a high rate of urogenital infection with C.trachomatis,with serotype E as the prevalent type.Since C.trachomatis serovar E-infected patients are likely to be missed by symptom-based screening and preventive strategies,standardized screening for and efficient treatment of urogenital C.trachomatis infection should be enhanced among low-tier FSWs for the prevention of C.trachomatis transmission.

Tests for the identification and isolation of Neis seria gonorrhoeae: a nationwide external quality assessment in 203 medical and healthcare facilities providing sexually transmitted disease services in 2011 / 中华皮肤科杂志

Objective To make a nationwide external quality assessment of tests for isolation and identification of N.gonorrhoeae in medical and healthcare facilities at different levels,and to analyze current problems.Methods Lyophilized quality control samples were uniformly delivered to 252 medical and healthcare facilities providing sexually transmitted disease (STD) services at different levels.Test results were analyzed by the National Center for STD Control,China Center for Disease Control and Prevention,and evaluation results were fed back to participating laboratories.Results Finally,test results were received from 203 (80.56％) facilities.The comprehensive score averaged at 87.14,and facilities achieving a comprehensive score of 80 or greater amounted to 80.30％ (163/203).The coincidence rate was 53.69％ (109/203) for all of the 5 quality control samples,82.76％ (168/203) and 87.68％ (178/203) respectively for two quality control samples containing only N.gonorrhoeae,86.21％ (175/203) and 96.06％ (195/203) respectively for a sample containing Neisseria sicca and a sample containing Enterococcus faecalis,69.46％ (141/203)for a sample containing different species of Neisseria.Conclusion The external quality assessment reveals a disparity in the capability to isolate and identify Neisseria among medical and healthcare facilities providing STD services at different levels.

Neisseria gonorrhoeae multi-antigen sequence typing using urine samples / 中华皮肤科杂志

Objective To determine Neisseria gonorrhoeae multi-antigen sequence typing (NG-MAST) sequence types in different geographical areas of China,including Changzhou and Yangzhou cities of Jiangsu province,Wuzhou and Hezhou cities in Guangxi Zhuang Autonomous region,Sanya and Qionghai cities of Hainan province,Jiangmen and Maoming cities of Guangdong province.Methods DNA was extracted using Qiagen DX extraction kits from 88 urine samples which were collected from male patients attending sexually transmitted disease (STD) clinics and positive for nucleic acid amplification test (NAAT) for N.gonorrhoeae.Two rounds of PCR were carried out to amplify the porB and tbpB genes of N.gonorrhoeae followed by gene sequencing.Sequence alignment was performed on the NG-MAST website (http://www.ng-mast.net) to determine the genotype of N.gonorrhoeae.Results The first-round PCR yielded positive results for porB and tbpB in 13.6％ (12/88) and 14.8％ (13/88),respectively,of these urine specimens,and 12 samples were successfully genotyped with the efficiency of genotyping being 13.6％.The amplification efficiency of second-round PCR was enhanced to 71.6％ and 72.7％ for porB and tbpB,respectively,and the efficiency of genotyping increased to 70.5％ (62/88).Compared with the first-round PCR,the second-round PCR showed an increase in amplification efficiency for porB and tbpB by 58.0％ and 57.9％ respectively,as well as in genotyping efficiency by 56.9％.Forty-five genotypes were identified in the 62 samples,including 40 known genotypes and 5 novel genotypes.Of these genotypes,ST1866 was the most abundant (6/62),followed by ST1972 (4/62) and ST3356 (4/62),all of which were from Jiangsu province.The ST532 genotype was identified in 3 samples from Guangdong province,ST2221 genotype in 2 samples from Guangxi Zhuang Autonomous region.Each of the remaining genotypes was identified in only 1 sample and scattered in all of these cities.The 5 novel MAST-genotypes were as follows:porB-892 and tbpB-46 (98％ similarity),porB-130 and tbpB-504 (96％ similarity),porB-2790 and tbpB-32 (99％ similarity),porB-1053 and tbpB-856 (99％ similarity).Conclusions Urine samples can be used for NG-MAST analysis,and two rounds of PCR can enhance the efficiency of genotyping.NG-MAST genotypes appear to be diverse in different geographical areas of China.

Hepatitis C virus infection among sexually transmitted disease clinic attendees infected with human immunodeficiency virus type 1: a survey in Guangxi Zhuang Autonomous Region / 中华皮肤科杂志

Objective To estimate the prevalence of hepatitis C virus (HCV) infection among sexually transmitted disease (STD) clinic attendees infected with human immunodeficiency virus type 1 (HIV-1) in Guangxi Zhuang Autonomous Region.Methods Totally,11 553 blood plasma samples were collected from STD clinic attendees in Guangxi Zhuang Autonomous Region,and subjected to HIV-1 antibody screening and confirmatory testing.Enzyme linked immunosorbent assay (ELISA) was performed to detect anti-HCV antibodies in 140 anti-HIV-1 antibody-positive samples and 282 anti-HIV-1 antibody-negative samples from age-and marital status-matched attendees.Chi-square test was performed to assess the differences in the prevalence rate of HCV infection between anti-HIV-1-negative and-positive samples,and Logistic regression analysis to evaluate the risk factors for HCV and HIV co-infection.Results The positivity rate of anti-HCV antibodies was 33.57％ (47/140)among anti-HIV-1-positive samples,significantly higher than that in anti-HIV-1-negative samples (1.06％ (3/282),x2 =94.66,P ＜ 0.05).Logistic regression analysis showed a statistical increase in the prevalence of HCV/HIV co-infection in individuals reporting more than one sexual partners compared with those reporting only one sexual partner (OR =2.4,95％ CI (1.0-5.6),P =0.05),and in intravenous drug users compared with non-intravenous drug users (OR =20.8,95％ CI(5.7-76.5),P ＜ 0.05).Conclusions HCV infection appears to be associated with HIV-1 infection,and comprehensive intervention on HIV-1-infected patients may slow down HCV transmission.

Development of a Taqman MGB real time fluorescence-based PCR assay for the detection of Mycoplasma genitalium / 中华皮肤科杂志

ObjectiveTo establish a rapid,sensitive and accurate method to detect Mycoplasma genitalium,and to evaluate the prevalence of M.genitalium among unlicensed prostitutes from Hezhou city in Guangxi Zhuang Autonomous Region.MethodsA pair of primers and Taqman MGB probe were designed and synthesized for the Pa gene of M.genitalium.Standard samples were prepared with the M.genitalium type strain G37.The established Taqman MGB real time fluorescence-based PCR assay was used to detect M.genitalium in the standard samples and cervical swab specimens collected from unlicensed prostitutes in Hezhou city of Guangxi Zhuang Autonomous Region.ResultsThe established Taqman MGB real time PCR exhibited a wide linear range( 1 × 10 copies/μl to 1 × 106 copies/μl,R2 =0.993),good repeatability(intra-assay variation;0.7％,inter-assay variation:1.09％) and hign sensitivity with the limit of detection being 10 copies/μl and limit of quantification being 50 copies/μl.As the assay showed,12.1％ of the 404 cervical swab samples were positive for M.genitalium.ConculsionThe Taqman MGB real time fluorescence-based PCR is a rapid and sensitive method for the quantitative and qualitative detection of M.genitalium.

Syphilis in outpatients at sexually transmitted disease clinics in Guangxi Zhuang Autonomous Region:an epidemiological study / 中华皮肤科杂志

ObjectiveTo investigate the prevalence of syphilis among outpatients in sexually transmitted disease (STD) clinics in Guangxi Zhnang Autonomous Region,and to assess the socioeconomic and behavioral factors associated with the infection.MethodsThe outpatients to 14 STD clinics in 8 cities of Guangxi Zhnang Autonomous Region were investigated with questionnaires by their doctors at the first visit.Venous blood samples were obtained from these outpatients and subjected to toludine red unheated serum test (TRUST) to screen for syphilis.Treponema pollidum particle agglutination (TPPA) was performed for TRUSTpositive samples.The epidemiological data were collected by using EpiData software,statistically analyzed by using SPSS13.0 software package.ResultsA total of 10 930 STD outpatients were recruited in the study,and 1297 samples were confirmed to be both TRUST and TPPA positive.The prevalence of syphilis was 11.9％ in all of the outpatients,14.3％ in female outpatients and 10.3％ in male outpatients,13.3％ in the outpatients of Zhuang nationality,and 11.4％ in those of Han nationalily.Multivariate analysis showed that syphilis was independently related to female sex[odds ratio(OR) 2.23,95％ confidence interval(CI) 1.69 - 3.00,P＜0.01 ],low educaiion level (middle school:OR 1.70,95％ CI 1.11 - 2.62,P ＜ 0.05; primary school or illiteracy,OR 1.98,95％ CI 1.13 - 3.46,P＜0.05),annual income of more than 30000 Yuan (OR 1.91,95％ CI 1.18 -3.10,P ＜ 0.01 ),commercial sex workers or having multiple sexual partners(OR 1.54,95％ CI 1.16 - 2.06,P ＜0.01 ).ConclusionsSyphilis serology should be the routine test in STD clinical settings in Guangxi region,and the intervention should be enhanced to control the prevalence of syphilis in high-risk populations.

Molecular typing of Treponema pallidum by analysis of three target genes / 中华皮肤科杂志

ObjectiveTo evaluate the performance of a three-gene typing system in the determination of Treponema pallidum (Tp) genotypes.MethodsTo determine the genotypes of Tp,three targets were assessed,including the number of 60 base-pair repeats,restriction fragment length polymorphism(RFLP) pattem of tprEGJ gene after MseI digestion and the sequence of tp0548 gene.The DNA extracted from the Nichols strain of Tp served as the positive control,and that from the moist ulcer of patients with genital herpes and negative RPR or TPPA test results served as the negative control.To validate the typing method,clinical specimens were collected from the moist skin lesions of patients with primary or secondary syphilis,and subjected to the amplification of polA gene by PCR.The enhanced molecular typing system was used to determine the genotypes of Tp in Tp DNA-positive specimens.ResultsThe Nichols strain harbored a genotype of 14a/a.No amplification of any of the three target genes was found in the negative control.The arp gene,tprEGJ gene and tp0548 gene were amplified from 94.1％,91.2％ and 94.1％ of the 40 clinical specimens,and the genotype was successfully determined by the three-gene typing system for 91.2％ of the clinical Tp strains.The predominant type of arp,tprEGJ and tp0548 genes was 14 repeats,d and f,respectively in these clinical Tp isolates.ConclusionThe enhanced molecular tying method for Tp exhibits high sensitivity,specificity and discrimination potential.