一种快速检测HIV抗体方法的评价

目的：探讨一种快速检测人类免疫缺陷病毒（HIV）抗体方法的敏感性和特异性,为艾滋病自愿咨询检测（VCT）及各种应急检测提供参考依据。方法：选择MSM调查人群血样1010份,平行用抗HIV-1／2快速检测（RT）试剂与抗HIV-1／2酶联免疫（ELISA）试剂进行检测,凡结果阳性者均用免疫印迹法（WB）进行确认。结果：710份血浆标本两种方法均有42份阳性,两者相符;300份全血标本用RT法检测,19份阳性,ELISA法22份阳性,3份不符标本后用血浆重复用RT法检测为阳性。所有阳性标本经WB确认均判为阳性。结论：HIV抗体快速检测试剂的使用需满足质量保证的要求并做好检测前后的咨询。建议使用经权威机构推荐的快速检测试剂;对于采供血机构,应使用ELISA法进行检测。     

Cost-effectiveness of adding a birth dose of hepatitis B vaccine in the Dafra district of the Hauts-Bassins Region in Burkina Faso (NéoVac Study)

BackgroundThe World Health Organization (WHO) recommends a first hepatitis B vaccine dose within 24 h of birth (HepB-BD) to prevent mother-to-child transmission. Evidence for this strategy’s economic value in Africa is limited. We assessed the costs and cost-effectiveness of adding HepB-BD to the current three-dose pentavalent schedule (HepB3) in the Dafra district of the Hauts-Bassins Region in Burkina Faso.MethodsUsing a decision tree combined with a Markov model, we estimated the expected number of life-years (LY) and disability-adjusted life-years (DALYs) saved, incremental costs, and incremental cost-effectiveness ratios (ICER) of HepB-BD + HepB3 versus HepB3 alone in Dafra’s 2017 birth cohort (n = 11,462). Institutional delivery rates, vaccine coverage, and vaccination costs from a health system perspective were estimated from field-collected data. We estimated the effectiveness of HepB-BD, age-specific transition probabilities, and horizontal transmission risks using data from previous African studies. Costs and health outcomes were discounted at an annual rate of 3%. We conducted one-way and probabilistic sensitivity analyses to assess uncertainty.ResultsIn the base-case analysis without discounting, HepB-BD + HepB3 yielded a net cost saving of US$18,979 and saved 163 DALYs compared with HepB3 alone. With discounting, HepB-BD + HepB3 compared with HepB3 resulted in an incremental cost of US$554 and 31 DALYs averted, translating into an ICER of US$18/DALY averted.In one-way sensitivity analyses, HepB-BD + HepB3 remained cost-effective (at the cost-effectiveness threshold of US$671 i.e. the Burkina Faso per-capita gross domestic product) for all parameter changes. However, results were very sensitive to variations in HepB-BD unit cost per vaccinated neonate and perinatal transmission risk in mothers carrying the hepatitis B e antigen. The probabilities of HepB-BD + HepB3 being cost-effective were 71.7% and 86.7%, at the cost-effectiveness thresholds of US$335 and US$671, respectively.ConclusionIntroducing HepB-BD in Burkina Faso is likely to be cost-effective.     

Evaluation of a panel of commercial kits for the detection of serum HIV-specific antibodies, and choice of alternative strategies for the serological diagnosis in Libreville (Gabon)

Nine commercially available kits for the screening of serum antibodies to the human immunodeficiency virus (HIV) were evaluated with a panel of 170 serum samples from adults in Gabon. The reference procedures showed that 96 samples had no antibodies, while 74 produced antibodies to HIV-1 (n=72) or HIV-2 (n=2). The sensitivity for 2 kits was less than 99% and the specificity of 3 less than 95%. Based on the panel of Gabonese serum samples, 4 of the 9 kits met the World Health Organization (WHO) acceptability criteria: sensitivity greater than 99% and specificity greater than 95%. Three kits available in Gabon that met these criteria were finally retained: Determine HIV-1/2, Genscreen Plus HIV Ag-Ab, and Immunocomb II HIV1&2 BiSpot. Of 18 configurations of alternative diagnosis strategies for HIV infection with these three kits, some WHO strategy II configurations (2 sequential assays) and all the WHO strategy III configurations (3 sequential assays) provided the maximum accuracy in diagnosing HIV infection in Gabon (sensitivity, specificity, positive predictive value, negative predictive value of 100%). The configuration using the Determine HIV-1/2 kit as first screening assay, followed by Genscreen Plus HIV Ag-Ab as a confirmatory assay, and finally, Immunocomb II HIV1&2 BiSpot as the third assay to discriminate samples discordant with the two first assays, was best, with high accuracy and less expense. The configuration of the two sequential rapid assays, Immunocomb II HIV1&2 BiSpot followed by Determine HIV-1/2, was also very accurate and reliable, as well as easiest to carry out (no need for ELISA technology), but it was twice as expensive as the first configuration with three sequential kits. In conclusion, when the laboratory facilities are available, the sequential diagnostic strategy of two rapid assays and a combined ELISA assay constitutes the best configuration, in terms of both accuracy and cost. When laboratory facilities are unavailable, sequential use of two rapid assays constitutes a convenient and accurate configuration, but is much more expensive.     

Meningitis serogroup W135 outbreak, Burkina Faso, 2002

In 2002, the largest epidemic of Neisseria meningitidis serogroup W135 occurred in Burkina Faso. The highest attack rate was in children <5 years of age. We describe cases from 1 district and evaluate the performance of the Pastorex test, which had good sensitivity (84%) and specificity (89%) compared with culture or PCR.     

免疫层析法与酶联免疫吸附法检测血中HBsAg、抗-HCV、抗-HIV的比较

〔目的〕分析比较乙型肝炎表面抗原(HBsAg)、丙型肝炎病毒抗体(抗-HCV)、HIV抗体(抗-HIV)的免疫层析快速试剂在实际应用中的灵敏度、特异度和准确度,为今后的工作提供参考。〔方法〕对502份健康体检血清标本同时用快速免疫层析法和酶联免疫吸附法(ELISA)检测HBsAg、抗-HCV、抗-HIV,对其灵敏度和特异度进行比较。〔结果〕与ELISA相比,502份血清标本中HBsAg金标法有2份假阴性,5份假阳性,灵敏度为99.6%,特异度为99.0%,准确度为98.6%;抗-HCV金标法无假阴性,有9份假阳性,灵敏度为100%,特异度为98.2%,准确度为98.2%;抗-HIV硒标法有2份假阳性,灵敏度为100%,特异度为99.6%,准确度为99.6%。〔结论〕金标法检测HBsAg灵敏度和特异度稍低于ELISA,有漏检和误检现象,而抗-HCV、抗-HIV快速法与ELISA比较灵敏度相当,而特异度稍低,因此,都要结合ELISA结果才可出具最终检测报告,抗-HIV的检测任何一种方法出现阳性时都要进行确认试验。     

A novel screening ELISA and a confirmatory Western blot useful for diagnosis and epidemiological studies of tularemia

A novel enzyme-linked immunosorbent assay (ELISA) and a confirmatory Western blot (WB) to detect human antibodies against Francisella tularensis were evaluated. The ELISA was based on partially purified lipopolysaccharide (LPS), the WB on whole antigen of F. tularensis. Positive WB showed a typical LPS ladder. Sensitivity and specificity of the ELISA, as assessed in 104 positive sera and 1149 'normal' sera from healthy young adults, were 99.0% and 97.1% respectively. Sensitivity of the WB was close to 100%, whereas specificity was 99.6%. Antibodies against the LPS of F. tularensis were detected in four of the 'normal' sera in both ELISA and WB. The assays were further evaluated using sera of individuals from Norway, Sweden and Kosovo suspected to be infected in tularemia outbreaks. Results revealed that the combination of ELISA and WB is suitable for laboratory confirmation of tularemia as well as for large-scale epidemiological studies.     

Effectively introducing a new meningococcal A conjugate vaccine in Africa: the Burkina Faso experience

A new Group A meningococcal (Men A) conjugate vaccine, MenAfriVac?, was prequalified by the World Health Organization (WHO) in June 2010. Because Burkina Faso has repeatedly suffered meningitis epidemics due to Group A Neisseria meningitidis special efforts were made to conduct a country-wide campaign with the new vaccine in late 2010 and before the onset of the next epidemic meningococcal disease season beginning in January 2011. In the ensuing five months (July-November 2010) the following challenges were successfully managed: (1) doing a large safety study and registering the new vaccine in Burkina Faso; (2) developing a comprehensive communication plan; (3) strengthening the surveillance system with particular attention to improving the capacity for real-time polymerase chain reaction (PCR) testing of spinal fluid specimens; (4) improving cold chain capacity and waste disposal; (5) developing and funding a sound campaign strategy; and (6) ensuring effective collaboration across all partners. Each of these issues required specific strategies that were managed through a WHO-led consortium that included all major partners (Ministry of Health/Burkina Faso, Serum Institute of India Ltd., UNICEF, Global Alliance for Vaccines and Immunization, Meningitis Vaccine Project, CDC/Atlanta, and the Norwegian Institute of Public Health/Oslo). Biweekly teleconferences that were led by WHO ensured that problems were identified in a timely fashion. The new meningococcal A conjugate vaccine was introduced on December 6, 2010, in a national ceremony led by His Excellency Blaise Compaore, the President of Burkina Faso. The ensuing 10-day national campaign was hugely successful, and over 11.4 million Burkinabes between the ages of 1 and 29 years (100% of target population) were vaccinated. African national immunization programs are capable of achieving very high coverage for a vaccine desired by the public, introduced in a well-organized campaign, and supported at the highest political level. The Burkina Faso success augurs well for further rollout of the Men A conjugate vaccine in meningitis belt countries.     

孕产妇HIV抗体筛查阳性与免疫印迹试验对比

目的探讨孕产妇人类免疫缺陷病毒（HIV）抗体筛查试验阳性与免疫印迹试验（WB）结果的关系。方法对33份孕产妇HIV抗体待复查样本,进行酶联免疫吸附试验（ELISA）及免疫印迹试验（WB）,对WB试验结果不确定者进行3个月、6个月随访检测,6个月随访样本补充HIV-1病毒载量检测,分析ELISA检测与WB试验检测结果。结果 33份样本ELISA复检均呈阳性,免疫印迹试验（WB）中20份确认为HIV-1抗体阳性,ELISA检测与WB试验阳性符合率为60.61%（20/33）;S/CO值〉8的20份样本,WB试验结果均为HIV-1抗体阳性,1〈S/CO值〈8的13份样本,WB试验5份为HIV抗体阴性,8份为HIV抗体不确定;6个月随访样本HIV-1病毒载量检测,结果均小于最低检测限（50 IU/ml）。结论孕产妇HIV抗体筛查（ELISA）假阳性较高,假阳性样本存在于WB试验抗体阴性和不确定结果之中;随着ELISA试验S/CO值增高,ELISA检测与WB试验的阳性符合率随之增高。     

四种艾滋病病毒抗体筛查试剂检测性能评价

目的 通过对比分析艾滋病病毒(HIV)抗体筛查阳性结果与免疫印迹试验(WB)结果,评价4种HIV抗体筛查试剂检测性能.方法 2004年1月至2009年6月,分别用中山生物工程有限公司、荷兰生物梅里埃有限公司、珠海丽珠有限公司生产的3种酶联免疫吸附试验(enzyme linked immunasorbent assay,ELISA)试剂初筛血清HIV抗体,用美国雅培Determine HIV-1/2胶体硒标试剂、原试剂复检.筛查阳性样本用WB法进行确认.结果 共检测206 151例患者血清HIV抗体,确认HIV抗体阳性193例(0.094%).3种ELISA试剂敏感度、阴性预期值均为100%;雅培试剂分别为93.93%、91.67%,其漏检的样本均为WB不确定.中山、梅里埃、丽珠、雅培试剂的特异度分别为99.88%、99.89%、99.96%、89.38%;阳性预期值(study predictive value of a positive test result,PVP)分别为35.58%、46.46%、76.61%、92.20%;功效分别为99.88%、99.89%、99.96%、91.98%;3种ELISA试剂ROC曲线下面积分别为0.93、0.99、0.95.丽珠的PVP明显高于中山(X~2=45.804,P=0.000)、梅里埃(X~2=25.231,P=0.000);梅里埃的PVP比中山高,但无统计学意义(X~2=2.488,P=0.115);雅培PVP最高(与丽珠相比,X~2=18.633,P=0.000).在WB确认阳性、不确定、阴性组,均存在S/CO值[样本(sample)吸光度值/临界值(cut off)]＜6或≥6的样本.中山试剂确认阳性组S/CO值(14.29±2.63)明显高于阳性-阴性组(2.80±3.25)(t=17.652,P=0.000).梅里埃试剂确认阳性组S/CO值(16.09±2.35)明显高于阳性-阴性组(2.14±1.91)(t=31.622,P=0.000).丽珠试剂确认阳性组S/CO值(11.54±1.95)明显高于阳性-不确定组(5.54±3.57)(t=6.386,P=0.000)、阳性-阴性组(3.25±2.41)(t=21.772,P=0.000);阳性-不确定组S/CO值则高于阳性-阴性组(t=2.301,P=0.033).结论 4种筛查试剂性能良好,根据S/CO值不能准确估计WB确认结果,筛查阳性后必须进行确认.     

三种乙型肝炎表面抗原检测方法在口岸体检中的应用评估

目的循证检验医学(EBLM)指导下,对电化学发光法(ECLIA)、酶联免疫吸附试验(ELISA)、胶体金免疫层析法等3种乙型肝炎表面抗原(HBsAg)检测方法进行评价,为口岸出入境人员寻求最佳的快速HBsAg检测方法和检测程序提供科学依据。方法ECLIA、ELISA、胶体金免疫层析法检测40份(HBsAg)标准血清盘标本,以血清盘预期结果为标准,比较3种方法灵敏度、特异度、总符合率。同时应用成本最小化分析方法,分别计算3种方法的成本。结果在血清盘HBsAg检测中,ECLIA法的灵敏度、特异性和总符合率皆达100.0%,Kappa值为1.000;而ELISA法国产科华试剂灵敏度76.9%、特异性100.0%和总符合率85.0%,Kappa值为0.432;胶体金免疫层析法灵敏度34.6%、特异性100.0%和总符合率57.5%,经Kappa检验其值为0.216。成本最小化分析结果显示:ECLIA法成本20元,ELISA法(科华,国产试剂)成本3元,胶体金免疫层析法成本3元,ECLIA法成本最高。结论ECLIA方法适用于需评估免疫状态,为接种疫苗做准备的出国留学人员、出国商务人员、入境留学生、入境商务人员的HBsAg检测。ELISA国产试剂适用于出国劳务人员、海员等一般人群。胶体金免疫层析法适用于紧急事情人员和日常复查的检测。     

一种HIV抗体化学发光诊断试剂的临床评价

目的临床评价北京科美东雅生物技术有限公司HIV抗体化学发光诊断试剂。方法用HIV抗体化学发光诊断试剂对既往已知经确证检测结果为HIV-1抗体阳性者标本500份和用酶联免疫吸附（ELISA）试剂检测为HIV-1／2抗体阴性者标本1500份的血清样品进行检测，并对检测结果进行比较。结果HIV抗体化学发光诊断试剂盒的敏感性、特异性和功效率均为100％。结论HIV抗体化学发光诊断试剂盒具有良好的应用价值。     

一种血液艾滋病病毒抗体免疫印迹试剂的现场评估

目的对Cambridge血液艾滋病病毒（HIV）-1抗体免疫印迹（WB）试剂进行现场评估并考核其在不同的HIV感染状态的人群中确认试验检测的敏感性和特异性.方法选取不同的现场,分别采集不同的HIV感染状态人群的血液样品,共计645份.经酶联免疫吸附试验（ELISA）检测后,分别使用Cambridge血液HIV-1抗体WB试剂盒和HIV blot 2.2 HIV-1/2型WB确认试剂盒进行平行对比试验.结果在已知既往HIV感染者中,Cambridge血液WB与Genelabs WB检测结果均为阳性,在此人群中上述2种确认试剂的敏感性均为100%.在398例HIV抗体ELISA检测为阴性的人群中,Cambridge血液WB试验23例为不确定;Genelabs血液WB试验86例为不确定,在此类人群中上述2种确认试剂的特异性分别为94.22%（375/398）和78.39%（312/398）.结论Cambridge血液HIV-1抗体WB试剂盒与Genelabs诊断公司的HIV blot 2.2 HIV-1/2型WB试剂盒的试验结果对比,在特异性方面前者优于后者.     

不同方法检测少数民族人群梅毒感染结果分析

目的探讨梅毒血清各种检测方法对梅毒的确诊及其治疗的灵敏性和特异性的临床指导意义。方法标本分别采用酶联免疫吸附试验、快速血清反应素环状卡片试验、荧光螺旋体抗体吸收试验、梅毒螺旋体血球凝集试验四种方法检测,并对结果进行比较分析。结果 ELISA、RPR、FTA-ABS、TPHA的敏感性分别为99.2%、98.3%、99.9%、91.3%,特异性为99.6%、86.4%、99.2%、94.2%。RPR与ELISA、FTA-ABS与TPHA2种方法检测结果经检验,差异无统计学意义(P>0.05),但RPR、ELISA与FTA-ABS、TPHA组间检测结果两两经检验,差异有统计学意义(P<0.05),且ELISA法的灵敏性和特异性均较好。在特异性方面,FTA-ABS法与ELISA、RPR、TPHA相比,差异有统计学意义(P<0.05),ELISA、RPR、TPHA两两间差异无统计学意义(P>0.05)。结论与FTA-ABS法的确认结果进行比较,RPR法不但有假阳性也有假阴性,存在梅毒漏检的可能;而ELISA法仅有假阳性、却无假阴性;ELISA法检出梅毒的能力强于RPR法。ELISA法是梅毒螺旋体血清学诊断试验的首选方法。近年来,ELISA法的应用研究主要趋向于联合使用两种或两种以上的重组梅毒螺旋体抗原,综合两种或多种抗原的不同特性,从而使ELISA法具有更高的特异性和敏感性。     

144例HIV抗体筛查阳性与免疫印迹试验结果对比研究

目的对比分析(ELISA方法检测)H IV抗体试验阳性与免疫印迹试验(WB)结果,探求H IV抗体筛查阳性与确证实验结果的关系。方法对144份ELISA方法检测阳性样本的结果与WB法确证结果进行比较,分析ELISA检测S/CO值变化与WB结果的关系。结果 144例H IV抗体筛查阳性标本经免疫印迹确证试验,阳性的有86例,均为H IV-1抗体阳性,阴性34例,结果不确定的24例。ELISA与WB的阳性符合率为59.7%。确认为阳性标本的S/CO平均值为15.7;不确定的标本S/CO值平均为3.04;阴性的标本S/CO值平均为1.24。在阳性标本中,p55条带出现的频率最低为48.8%,其它均达到85%以上,其中gp160、gp120达到100%;不确定标本中,p24条带出现的几率最高,达75%,其它都在20%以下。结论筛查试验存在假阳性结果,H IV抗体结果的报告必须以确认结果为准。高S/CO值预示H IV抗体阳性的可能性较大,而S/CO值较低时H IV抗体阴性及不确定的可能性较大。WB确认方法在不确定标本检测中存在一定的缺陷。     

143例HIV初筛阳性标本免疫印迹试验结果分析

目的 使用酶联免疫吸附法(ELISA)、硒标免疫层析法(Determine)及免疫印迹法(WB)对143例血清标本做初筛及确认试验。方法 操作及结果判断按照《全国艾滋病检测技术规范》施行。结果 143例初筛试验阳性经免疫印迹试验确认141例HIV—1阳性，且条带出现百分率gp^120、p^66为100％，gp^160、gp^41、p^51、p^34、p^55为90％以上，1例HIV—1合并gp^36出现。结论 初筛结果有假阳性须经确认试验才能得出准确结果。     

蛋白印迹试验和第4代酶联免疫吸附试验艾滋病病毒检测结果

目的 对2011—2014年艾滋病病毒(HIV)抗体筛查阳性标本的确证试验结果进行分析。方法 2011—2014年收集天津市HIV高危人群的实验室资料,采用第4代酶联免疫吸附试验(ELISA)及胶体硒试验,对送检的和本筛查中心实验室检出的220例筛查阳性标本,使用蛋白印迹试验(WB)进行确证试验。结果 220例阳性标本,确证结果为阳性的病例180例(81.82%),经过1周,进行2次确证试验,阳性符合率为180例(100%)。阴性病例共10例(4.55%),不确定病例共30例(13.64%)。确证的180例中,gp160、gp120出现率达到100%,p55、p39阳性率为66%(119例)、61%(110例),gp41出现率为96%(173例),p51出现率为93%(167例),p31出现率为(160例),p24出现率为98%(176例)。结论 HIV抗体初筛试验结果存在假阳性,WB和第4代ELISA筛查试验可提高对艾滋病病毒的检查准确率,对WB试验中不确定标本需做好随访工作,避免艾滋病患者流失。     

血液唾液和尿液HIV抗体快速检测及免疫印迹试剂评估

目的对Calypte公司血液、唾液和尿液3种HIV-1／2抗体快速检测试剂与尿液和血液两种HIV-1抗体免疫印迹试剂进行评估,为引进检测性能良好的试剂提供参考。方法平行采集84位HIV阳性感染者指尖血、唾液、尿液和静脉血标本,分别用Calypte公司的3种快速试剂进行HIV-1／2抗体检测,并与梅里埃血液ELISA参比试剂检测结果比较;尿液和静脉血标本分别用Calypte公司的两种免疫印迹试剂进行检测,并与GerieLabs公司的血液免疫印迹参比试剂检测结果进行比较。结果血液、唾液和尿液快速检测试剂的敏感性分别为100％、100％和96．42％,3种快速试剂检测结果与梅里埃血液ELISA检测结果一致性分别为100％、100％和96．42％;血液和尿液HIV-1WB试剂检测结果与GeneLabs参比试剂的一致性为100％。结论Calypte公司3种HIV-1／2抗体快速检测试剂和两种HIV-1抗体免疫印迹试剂与相应的参比试剂有很好的一致性。     

Detection of meningitis epidemics in Africa: a population-based analysis.

Portions of sub-Saharan Africa are subject to major epidemics of meningococcal meningitis that require early detection and rapid control. We evaluated the usefulness of weekly meningitis rates derived from active surveillance data in Burkina Faso for detecting a meningitis epidemic. By analysing the rates of disease in 40 x 40km2 areas within a study region of Burkina Faso, we found that a threshold of 15 cases/100,000/week averaged over 2 weeks was 72-93% sensitive and 92-100% specific in detecting epidemics exceeding 100 cases/100,000/year. During epidemic periods, the positive predictive value of this threshold approached 100% for detecting local epidemics. Additionally, meningitis incidence was proportional to village size, with villages greater than 8000 having the highest disease rates during a major group A meningococcal epidemic in 1983-1984. Despite the rudimentary nature of surveillance data available in many developing countries, these data can be used to detect the early emergence of meningitis epidemics. Additional studies are needed to determine the relevance of this approach for detecting epidemics.     

间接免疫荧光试验在HIV抗体检测中的应用

目的：评价间接免疫荧光试验（IFA）作为一种确认实验在HIV抗体检测中的应用。方法：用T嗜性的HIV毒株感染MT4细胞制备抗原片,并与不同稀释度的免疫血清相结合,再滴加荧光素标记的羊抗人IgG,在荧光显微镜下检测特异性免疫荧光,结果：用IFA检测43分血清样本（其中包括17份WB阳性血清,14份阴性血清和12份ELISA阳性或WB出现可疑条带但按标准不能判为阳性的可疑血清）,除HIV－2血清呈阴性反应外,其余检测结果与WB相符,IFA对HIV－1抗体检测的敏感性和特异性均达到100％,HIV－1阳性血清抗体最高滴度达到1：10240。结论：IFA可用于HIV－1抗体检测的确认,它可作为Western Blot的一种补充或替代实验。     

广州市发现首例HIV-1/HIV-2混合感染

目的探讨首例HIV-1/HIV-2混合感染发现的经过以及对本地艾滋病预防控制的意义。方法对广州市某医院一名门诊患者血样2份(包括初筛试验前后各1份)进行HIV抗体检测。初筛试验采用酶联免疫吸附试验(ELISA)和明胶颗粒凝集试验(PA),确认试验采用蛋白印迹法(WB)。结果患者为男性,非洲马里人,血清经初筛试验为HIV阳性。初筛阳性的血清,以HIV-1/HIV-2 HIVBLOT2.2试剂做蛋白印迹试验(WB),确认为HIV-1阳性,同时HIV-2的特异条带gp36阳性;样本再用HIV-2型的BLOT1.2膜条确认为HIV-2阳性。结论被检者为HIV-1/HIV-2混合感染,这是广州市、广东省首次发现HIV-1/HIV-2混合感染,提示HIV-2可能会以对外交流等方式传入广东。