Gingivocrevicular transudate for HIV screening

588 Clinical Mic obiology and In ec ion, Volume 3 Numbe 5, Oc obe 1997 ha he co ec ional acili ies, which a p esen ha e inadequa e in ec ion con ol p ac ices and isola ion acili ies o p e en ansmission o M. ube culosis, ully comply wi h he cu en guidelines o educe he isk o ube culosis sp ead [14,15]. Acknowledgmen s This wo k was pa ly suppo ed by he Comisi6n In e minis e ial de Ciencia y Tecnologia (CICYT) g an SAF95/0435 by he Minis e io de Educaci6n y Ciencia. Jauie Azna I, Hassan S&*, Mada C. Conejo’, Jos6 C. Paloma es’ ‘Dp o Mic obiologia, Apdo 914, Uni e sidad de Se illa, 41080-Se illa, Spain; *Depa men o Mic obiology, Uni e si y o Se ille, Se ille, Spain Re ised e sion accep ed 3 May 1997 Re e ences 1. G ei inge RB, Kee us C, G abau J, e al. T ansmission o mul id ug- esis an ube culosis among immunocom- p omised pe sons in a co ec ional sys em. New Yo k 1991. 2. Sepkowi z KA, R&ah J, Riley L, Kiehn TE, Ams ong D. Tube culosis in he AIDS e a. Clin Mic obiol Re 1995; 8: MMWR 1992; 41: 507-9. 180-99. 3. Valway SE, Richa ds SB, Ko aco ich J, G ei inge RB, C aw o d JT, Dooley SW. Ou b eak o mul i-d ug- esis an ube culosis in a New Yo k S a e P ison 1991. Am J Epidemiol 1994; 140: 113-22. 4. De Ma ch P. Si uaci6n ac ual de la ube culosis en Espaiia. Med Clin (Ba c) 1991; 97: 463-72. 5. Ma in V, Gonzalez P, Cayla JA, e al. Case- inding o pulmona y ube culosis on admission o a peni en ia y cen e. Tube cle Lung Dis 1994; 75: 49-53. 6. He mans PWM, an Soolingen D, Dale JW, e al. Inse ion elemen IS986 om Mycobac e ium ube culosis: a use ul ool o diagnosis and epidemiology o ube culosis. J Clin Mic obiol 1990; 28: 2051-8. 7. Small PM, Sha e RW, Hopewell PC, e al. Exogenous ein ec ion wi h mul id ug- esis an Mycobac e ium ube culosis in pa ien s wi h ad anced HIV in ec ion. N Engl J Med 8. an Soolingen D, He mans PW, de Haas PEW, Soll DR, an Embden JDA. Occu ence and s abili y o inse ion sequences in Mycobac e ium ube culosis complex s ains: e alua ion o an inse ion sequence-dependen DNA poly- mo phism as a ool in he epidemiology o ube culosis. J Clin Mic obiol 1991; 29: 2578-86. 9. Kubica GPW, Dye E, Cohn ML, Middleb ook DH. Spu um diges ion and decon amina ion wi h N-ace yl-L-cys eine- 1993; 328: 1137-44. sodium hyd oxide o cul u e o mycobac e ia. Am Re Respi Dis 1963; 87: 775-9. 10. an Embden JDA, Donald Ca e M, C aw o dJT, e al. S ain iden i ica ion o Mycobac e ium ube culosis by DNA inge - p in ing: ecommenda ions o a s anda hzed me hodology. J Clin Mic obiol 1993; 31(2): 406-9. 11. Azna J, Sa i H, Rome o J, Alejo A, G acia MD, Paloma es JC. Nosocomial ansmission o ube culosis in ec ion in pedia ic wa ds. Peha In ec Dis J 1995; 14: 44-8. 12. He mans PWM, Messadi F, Gueb exabbe H, e al. Analysis o he popula ion s uc u e o Mycobac e ium ube culosis in E hiopia, Tunisia, and he Ne he lands: use ulness o DNA yping o global ube culosis epidemiology. J In ec Dis 1995; 171: 1504-13. 13. Ba nes PE El-Haj H, P es on-Ma in S, e al. T ansmission o ube culosis among he u ban homeless. JAMA 1996; 275: 14. The ole o BCG accine in he p e en ion and con ol o ube culosis in he Uni ed S a es. A join s a emen by he Ad iso y Commi ee o Elimina ion o Tube culosis and he Ad iso y Commi ee on Immuniza ion P ac ices. 15. P e en ion and con ol o ube culosis in co ec ional acili ies. Recommenda ions o he elimina ion o ube - culosis. MMWR 1996; 45: 1-27. 305-7. MMWR 1996; 45: 1-14. Gingi oc e icula ansuda e o HIV sc eening Clin Mic obiol In ec 1997; 3: 588-590 The use o sali a as an al e na i e biological luid o se um in diagnosis o sc eening o in ec ious diseases by an ibody de ec ion has been he main subjec o se e al a icles in he las ew yea s [l-61. Sah a samples can be ob ained simply, wi hou specialized pe sonnel, and he p ocess is non- auma ic o he pa ien and economic and poses no con amina ion isks o heal h wo ke s. Fo hese easons, sali a samples may be o g ea u ili y in unde de eloped na ions, whe e he e is a se e e sho age o pe sonnel and specialized equipmen . Sali a is a mix u e o he sec e ions p oduced by he sali a y glands and gingi al c e icula ansuda e (GCT). The use o GCT, which has a g ea e concen a ion o immunoglobulins (Ig) o he IgG ype han does comple e sali a [7], seems o imp o e de ec ion o such Ig in he samples [8]. An o al de ice wi hou p ese a i es (Sali e e, S a s ed , Leices e , UK) was used o ob ain he GCT samples. The de ice is a cylinde o co on wool con- ained in a polys y ene double ube wi h a snap op (p ice USA $0.20). The co on wool was placed in he la e al gingi al old un il he indi idual could pe cei e ha i had become so because o i s abso p ion o GCT. The samples hus ob ained we e cen i uged a 4000 g o 15 min, p oducing a supe na an which was Sho Communica ions and Le e s o he Edi o s 589 aliquo ed and ozen a -80°C un il p ocessing. A se um sample was simul aneously ob ained by ene- punc u e om each o he indi iduals included in he s udy. These samples we e hen aliquo ed and s o ed a -80°C un il p ocessed. A o al o 183 pai ed samples o GCT and se um we e analyzed (50 CDC s age 11, 10 CDC s age 111, 33 CDC s age I 26 isk ac o indi iduals and 64 blood dono s). In bo h ypes o sample, he p esence o IgG agains HIV-I and HIV-2 was de ec ed. Wi h GCT samples, we used a new do blo (Mul ispo HIV- 1 /HIV-2, Sano i Diagnos ics Pas eu , F ance) capable o disc imina ing be ween bo h IgG HIV-1 and IgG HIV-2. Fo he sc eening o se um samples, an indi ec enzyme immunoassay (EIA) (VIDAS HIV 1+2, bioMk ieux, F ance) was used. All eac i e samples we e con i med by Wes e n blo (WB) (Bioblo HIV- I plus, Genelabs Diagnos ics, Swi ze land). The WB used combines speci ic p o eins o HIV-1 and a syn he ic pep ide (gp36) o HIV-2 in he same s ip; WHO c i e ia [9] we e used o con i ma ion o sc eening eac i i y. The esul s p esen ed in Table 1 show ha all GCT samples non- eac i e by do blo belonged o non- in ec ed indi iduals (EIA nega i e), excep o one sample om a pa ien o he CDC s age IV g oup, which was nega i e by do blo bu posi i e by GCT WB ( eac i i y agains gp160, gp120 and gp41). The GCT samples eac i e by do blo co espond o in ec ed indi iduals, wi h he excep ion o one sample om a non-in ec ed indi idual also nega i e by GCT WB. The cha ac e is ics o his es in compa ison wi h se um WB esul s ha e been as ollows: 99% sensi i i y and 99% speci ici y (posi i e p edic i e alue (PPV) and nega i e p edic i e alue (NPV) we e 0.9). Se e al easons may be sugges ed in he case o he non- eac i e sample om he indi idual belonging o CDC s age IV The i s is he immunologic de e io a- ion p esen ed by indi iduals in he inal s ages o he in ec ion, yielding a lowe IgG se um concen a ion and e en loss o conc e e an ibodies (ie. p24 an i- bodies); second, GCT may he e o e no p esen eac i i y when analyzed by insu icien ly sensi i e echniques. The use o a mo e sensi i e es (WB) leads in his case o he de ec ion o IgG agains en elope p o eins in he GCT. The con i med p esence o HIV- 1 en elope an ibody in GCT is highly sugges i e o HIV-1 in ec ion and also co ela es wi h he eac i i ies ob ained in he se um WB o his indi idual. The sali a y de ice used p esen s wo main ad an ages. The i s and mos impo an is ha i can ob ain an adequa e olume (= 1 ml) o sample analysis by di e en echniques and s o age. Second, in con as o he case wi h o he de ices, IgG sample concen- a ion does no a y so much, because he sample is no dilu ed. The e is no expe imen al e idence ha he concen a ion o Ig in GCT declines du ing s o age, bu any such changes would be minimal, because he samples we e no dilu ed in any p ese a i e bu e and hey we e ozen a e eco e y, hus minimizing he ac ion o sali a enzymes. GCT samples (USA $0.20) a e cheape o ob ain han se um samples (USA $0.60 o sy inge +needle+ ube). Fu he mo e, he GCT p o- cess pe mi s a single indi idual o collec samples om se e al pa ien s a once, ins ead o he indi idualized sample collec ion equi ed wi h se um. Ou esul s con i m p e ious wo k by o he s [10,11] bu using a simple de ice o sampling and es ing. The use o GCT ins ead o sali a has been p oposed in se e al a icles [8,10,11] o acili a e he de ec ion o HIV IgG due o he inc eased amoun o his ype o Ig. In conclusion, he use o do blo oge he wi h he o al GCT de ice could o e an al e na i e o sc eening o in ec ion p oduced by HIV in wide popula ion g oups, because o i s low cos and easy applica ion. Fu he s udies should be pe o med in pa ien s wi h ad anced s ages o he disease o de e - mine i he lloss in sensi i i y could be due o he immunologic condi ion in his pa icula s age o he disease. Table 1 De ec ion o HIV-1 an ibodies in he se um and GCT o di e en popula ion g oups G oup No. Do cc EIAsc unl mp" S age I1 50 50 50 50 S age I11 10 10 10 10 S age IV 33 32 33 33 Subjec s a isk o HIV in ec ion 26 0 0 0 Blood dono s 64 lb 0 0 Re e ences P udencio Ma inez, josk Ma ia Ei os Raul O iz dc Leja a u An onio Rod kuez To es Se icio de Mic obiologia Clinica, Hospi al Uni e si a io, Valladolid, Spain Accep ed 25 Ap il 1997 To al 183 93 93 93 1. Pa IV. Pe m KR. Mo ime PP. Sensi i e assa s o i al ,_l, , , an ibodies in sali a: an al e na i e o ec 011 ce um. Lance "Samples wi h posi i e Wes e n blo in se um and GCT. "Sample wi h nega i e Wes e n blo in GCT. 1987; 2: 72-5. 590 Clinical Mic obiology and In ec ion, Volume 3 Numbe 5, Oc obe 1997 4. 5. 6. 7. 8. 9. 10. 11. 2. Pa y JV, Pe y KR, Panday S, Mo ime PP. Diagnosis o hepa i is A and B by es ing sali a. J Med Vi ol 1989; 28: 255-60. 3. Thieme T, Yoshiha a P, Piacen ini S, Belle M: Clinical e alua ion o o al luid samples o diagnosis o i al hepa i is. J Clin Mic obiol 1992; 30: 1076-9. Luch E, Albe J, Linde A, e al. Human ihmunode iciency i us ype l and cy omegalo i us in sali a. J Med Vi ol 1993; 39. 15Gh2 The HBe Ag p ecu so is encoded by he 637-bp p eco e/co e open eading ame s a ing a nucleo ide 1814 o he HBV minus s and genome (nomencla u e acco ding o Galibe e al [4]), The equen ly desc ibed mu a ion is a G o A subs i u ion a he 3’ end o he p e-C gene (nucleo ide 1896), which con e s codon 28 o yp ophan (TGG) o a e mina- ion codon (TAG) [S], esul ing in he ailu e o HBe - . . . - - .. - . F e ichs R, H oo i M, Eskes N, Soe L. Compa ison o sali a and se um o HIV su eillance in de eloping coun ies. Lance 1992; 340: 1496-9. Akke R, Hoek J, Akke W, e al. De ec ion o HIV an ibodies in sali a as a ool o epidemiological s udies. Pa y JV. A specimen o con enience. Diagn Vi ol 1991; 3: Ma inez P, O iz de Leja azu K, Ei os JM, e al. Compa ison o wo assays o de ec ion o HIV an ibodm in sali a. Eu J Clin Mic obiol 1995; 14: 330-6. Wo ld Heal h O ganiza ion. AIDS. P oposed WHO c i e ia o in e p e ing esul s iom Wes e n blo assays o HIV-1, HIV-2 and HTLV-I/HTLV-11. Wkly Epidemiol Rec 1990; G6mez C, Gu i ez M, Ma inez-Acacio P, So iano V. E alua ion o a new sali a collec ion de ice o HIV an i- body sc eening pu poses. Vox Sang 1994; 66: 244. Gallo D, Geo ge R, Fi chen J, Golds ein A, Hindahl M. E alua ion o a sys em using o al mucosal ansuda e o HIV-1 an ibody sc eening and con i ma o y es ing. JAMA 1997; 277: 254-8. AIDS 1992; 6: 953-7. 13-15. 65: 281-3. Mo e on HBe Ag-nega i e mu an s in ch onic HBV in ec ion Clin Mic obiol In ec 1997; 3: 590-591 Ch onic in ec ion wi h hepa i is B i us (HBV) leads o a wide spec um o li e diseases, including asymp- oma ic ca ie s a e, ch onic ac i e hepa i is, ci hosis and hepa ocellula ca cinoma. Pa ien s p esen ing wi h ch onic ac i e hepa i is ypically ha e e idence o HBV eplica ion ma ked by high le els o HBV DNA, HBV su ace an igen (HBs Ag) and he e an igen (HBe Ag) in hei se um [l]. In la e s ages o he disease, se ocon e sion om HBe Ag o an i-HBe an ibody classically indica es a ansi ion o a ‘non- eplica i e’ phase, in which nec o-in lamma o y ac i i y ceases. In some cases, howe e , an i-HBe se ocon e sion is no ollowed by such a a o able ou come, and ma ke s o HBV eplica ion emain de ec able [2]. Ca man e a1 [3] ha e s udied such pa ien s and ound ha hey ca ied HBV mu an s which canno syn hesize he p eco e p ecu so p o ein and he e o e HBe Ag. Ag p oduc ion and he cessa ion o i s sec e ion in o he bloods eam. I is o en associa ed wi h a G o A silen mu a ion a nucleo ide (n ) posi ion 1899. The equency o he n 1896 mu a ion a ies in di e en s udies, anging om 46% [6] o 80% [3]. This a i- abili y led us o in es iga e he equency o he n 1896 mu a ion and he p esence o o he pu a i e mu a ions in a coho o F ench pa ien s. We analyzed 32 se um samples om ch onically in ec ed pa ien s showing disco dan se ologic p o iles, i.e. posi i e o HBs Ag and HBV DNA, nega i e o HBe Ag and posi i e o an i-HBe an ibody. Se um samples we e submi ed o hea ea men (100 “C) o 30 min and hen cla i ied by cen i uga ion (12000g) o 30 min. This apid echnique o HBV DNA p epa a ion had been alida ed in p elimina y s udies as compa ed ei he o classical p o einase K-sodium dodecylsul a e (SDS) lysis ollowed by phenol ex ac ion o o NaOH ea men o he se um. We applied an ampli ica ion-c ea ed es ic ion si e me hod de i ed om ha desc ibed by Lindh e a1 [7] o de ec ing he n 1896 G o A mu a ion. Acco ding o his p ocedu e, he n 1865-2058 egion o HBV DNA was ampli ied using P1 p ime wi h a misma ch a posi ion 1893. When he n 1896 G o A mu a ion was p esen , he 194-bp PCR p oduc exhibi ed a BSU 361 es ic ion si e which could be shown by enzyme diges ion and aga ose gel elec opho esis. P io o hges ion, a plasmid con aining a unique BSU 361 es ic ion si e was added as a con ol o he comple ion o diges ion. The speci ici y o he PCR eac ion was checked by hyb idiza ion wi h a digoxigenin-labeled p obe speci ic o he HBV p eco e egion. In pa allel, he n 1728-2160 egion o HBV DNA was ampli ied o he de e mina ion o nucleo ide sequence using a T7 polyme ase sequencing ki (Pha macia). A e diges ion wi h BSU 361, we obse ed h ee pa e ns o mig a ion in an aga ose gel among he 32 samples es ed. Fi e cases (16%) showed a diges ed p oduc o 165 bp cha ac e is ic o he G o A mu a ion. Twen y cases (62%) showed an undiges ed PCR p o- duc o 194 bp. In se en (22%) cases, wo agmen s comig a ed in he aga ose gel, one o 194 bp, and one o 165 bp, co esponding o a mixed i al popula ion (wild ype and mu an ). In hese cases, he possibili y o pa ial diges ion wi h BSU 361 was uled ou by